Spirocyclic compounds as modulators of chemokine receptor activity

ABSTRACT

The present application describes modulators of chemokine receptor activity of formula I 
     
       
         
         
             
             
         
       
     
     or stereoisomers or pharmaceutically acceptable salts thereof. In addition, methods of treating and preventing inflammatory diseases such as asthma and allergic diseases, as well as autoimmune pathologies such as rheumatoid arthritis and transplant rejection using modulators of formula (I) are disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Divisional of U.S. patent application Ser. No.13/500,323, filed on Apr. 5, 2012, which is a U.S. national phase ofInternational Application No. PCT/US2010/051577, filed on Oct. 6, 2010,which claims priority of U.S. Ser. No. 61/249,364, filed Oct. 7, 2009,incorporated by reference herein in their entirety.

FIELD OF THE INVENTION

This invention relates generally to spirocyclic compounds as modulatorsof chemokine receptor activity, pharmaceutical compositions containingthe same, and methods of using the same as an agent for treatment andprevention of inflammatory diseases, allergic and autoimmune diseases,and in particular, rheumatoid arthritis and transplant rejection.

BACKGROUND OF THE INVENTION

Chemokines are chemotactic cytokines, of molecular weight 6-15 kDa, thatare released by a wide variety of cells to attract and activate, amongother cell types, monocytes, macrophages, T and B lymphocytes,eosinophils, basophils and neutrophils. There are two major classes ofchemokines, CXC and CC, depending on whether the first two cysteines inthe amino acid sequence are separated by a single amino acid (CXC) orare adjacent (CC). The CXC chemokines, such as interleukin-8 (IL-8),neutrophil-activating protein-2 (NAP-2) and melanoma growth stimulatoryactivity protein (MGSA) are chemotactic primarily for neutrophils and Tlymphocytes, whereas the CC chemokines, such as RANTES, MIP-1α, MIP-1β,the monocyte chemotactic proteins (MCP-1, MCP-2, MCP-3, MCP-4, andMCP-5) and the eotaxins (-1 and -2) are chemotactic for, among othercell types, macrophages, T lymphocytes, eosinophils, dendritic cells,and basophils.

The chemokines bind to specific cell-surface receptors belonging to thefamily of G-protein-coupled seven-transmembrane-domain proteins whichare termed “chemokine receptors.” On binding their cognate ligands,chemokine receptors transduce an intracellular signal though theassociated trimeric G proteins, resulting in, among other responses, arapid increase in intracellular calcium concentration, changes in cellshape, increased expression of cellular adhesion molecules,degranulation, and promotion of cell migration. There are at least tenhuman chemokine receptors that bind or respond to CC chemokines with thefollowing characteristic patterns: CCR-1 (or “CKR-1” or “CC-CKR-1”)[MIP-1α, MCP-3, MCP-4, RANTES] CCR-2A and CCR-2B (or “CKR-2A”/“CKR-2B”or “CC-CKR-2A”/“CC-CKR-2B”) [MCP-1, MCP-2, MCP-3, MCP-4, MCP-5];CCR-3(or “CKR-3” or “CC-CKR-3”) [eotaxin-1, eotaxin-2, RANTES, MCP-3, MCP-4];CCR-4 (or “CKR-4” or “CC-CKR-4”) [TARC, MDC]; CCR-5 (or “CKR-5” OR“CC-CKR-5”) [MIP-1α, RANTES, MIP-113]; CCR-6 (or “CKR-6” or “CC-CKR-6”)[LARC]; CCR-7 (or “CKR-7” or “CC-CKR-7”) [ELC]; CCR-8 (or “CKR-8” or“CC-CKR-8”) [1-309]; CCR-10 (or “CKR-10” or “CC-CKR-10”) [MCP-1, MCP-3];and CCR-11 [MCP-1, MCP-2, and MCP-4].

In addition to the mammalian chemokine receptors, mammaliancytomegaloviruses, herpesviruses and poxviruses have been shown toexpress, in infected cells, proteins with the binding properties ofchemokine receptors. Human CC chemokines, such as RANTES and MCP-3, cancause rapid mobilization of calcium via these virally encoded receptors.Receptor expression may be permissive for infection by allowing for thesubversion of normal immune system surveillance and response toinfection. Additionally, human chemokine receptors, such as CXCR4, CCR2,CCR3, CCR5 and CCR8, can act as co-receptors for the infection ofmammalian cells by microbes as with, for example, the humanimmunodeficiency viruses (HIV).

The chemokines and their cognate receptors have been implicated as beingimportant mediators of inflammatory, infectious, and immunoregulatorydisorders and diseases, including asthma and allergic diseases, as wellas autoimmune pathologies such as rheumatoid arthritis andarthrosclerosis (reviewed in: Carter, P. H., Current Opinion in ChemicalBiology 2002, 6, 510; Trivedi et al., Ann. Reports Med. Chem. 2000, 35,191; Saunders et al., Drug Disc. Today 1999, 4, 80; Premack et al.,Nature Medicine 1996, 2, 1174). For example, the chemokine macrophageinflammatory protein-1 (MIP-1α) and its receptor CC Chemokine Receptor 1(CCR-1) play a pivotal role in attracting leukocytes to sites ofinflammation and in subsequently activating these cells. When thechemokine MIP-1α binds to CCR-1, it induces a rapid increase inintracellular calcium concentration, increased expression of cellularadhesion molecules, cellular degranulation, and the promotion ofleukocyte migration.

In addition, demonstration of the chemotactic properties of MIP-1α inhumans has been provided experimentally. Human subjects, when injectedintradermally with MIP-1α, experienced a rapid and significant influx ofleukocytes to the site of injection (Brummet, M. E., J. Immun. 2000,164, 3392-3401).

Demonstration of the importance of the MIP-1α/CCR-1 interaction has beenprovided by experiments with genetically modified mice. MIP-1α −/− micehad normal numbers of leukocytes, but were unable to recruit monocytesinto sites of viral inflammation after immune challenge. Recently,MIP-1α −/− mice were shown to be resistant to collagen antibody inducedarthritis. Likewise, CCR-1 −/− mice were unable to recruit neutrophilswhen challenged with MIP-1α in vivo; moreover, the peripheral bloodneutrophils of CCR-1 null mice did not migrate in response to MIP-1α,thereby demonstrating the specificity of the MIP-1α/CCR-1 interaction.The viability and generally normal health of the MIP-1α −/− and CCR-1−/− animals is noteworthy, in that disruption of the MIP-1α/CCR-1interaction does not induce physiological crisis. Taken together, thesedata lead one to the conclusion that molecules that block the actions ofMIP-1α would be useful in treating a number of inflammatory andautoimmune disorders. This hypothesis has now been validated in a numberof different animal disease models, as described below.

It is known that MIP-1α is elevated in the synovial fluid and blood ofpatients with rheumatoid arthritis. Moreover, several studies havedemonstrated the potential therapeutic value of antagonism of theMIP-1α/CCR1 interaction in treating rheumatoid arthritis.

It should also be noted that CCR-1 is also the receptor for thechemokines RANTES, MCP-3, HCC-1, Lkn-1/HCC-2, HCC-4, and MPIF-1 (Carter,P. H., Curr. Opin Chem. Bio. 2002, 6, 510-525). Since it is presumedthat the new compound of formula (I) described herein antagonizes MIP-1αby binding to the CCR-1 receptor, it may be that this compound is alsoan effective antagonist of the actions of the aforementioned ligand thatare mediated by CCR-1. Accordingly, when reference is made herein to“antagonism of MIP-1α,” it is to be assumed that this is equivalent to“antagonism of chemokine stimulation of CCR-1.”

Recently, a number of groups have described the development of smallmolecule antagonists of MIP-1α (reviewed in: Carson, K. G. et al., Ann.Reports Med. Chem. 2004, 39, 149-158).

SUMMARY OF THE INVENTION

Accordingly, the present invention provides novel antagonists or partialagonists/antagonists of MIP-1α or CCR-1 receptor activity of the formula

or stereoisomers or pharmaceutically acceptable salts thereof.

The present invention provides pharmaceutical compositions comprising apharmaceutically acceptable carrier and a therapeutically effectiveamount of at least one of the compounds of the present invention or apharmaceutically acceptable salt form thereof.

The present invention provides a method for treating rheumatoidarthritis and transplant rejection, comprising administering to a hostin need of such treatment a therapeutically effective amount of at leastone of the compounds of the present invention or a pharmaceuticallyacceptable salt form thereof.

The present invention provides a method for treating inflammatorydiseases, comprising administering to a host in need of such treatment atherapeutically effective amount of at least one of the compounds of thepresent invention or a pharmaceutically acceptable salt form thereof.

The present invention provides novel spirocyclic compounds for use intherapy.

The present invention provides the use of novel spirocyclic compoundsfor the manufacture of a medicament for the treatment of inflammatorydiseases.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

In one embodiment, the present invention provides novel compounds offormula (I):

or stereoisomers or pharmaceutically acceptable salt forms thereof,wherein:

-   -   the dashed line represents an optional double bond;    -   Ring A is an optionally substituted three to nine membered mono-        or bicyclic heterocyclic ring;    -   T is

-   -   W is —OH;    -   Z is a O or S;    -   R and R₁ are independently hydrogen, alkyl, halo, C═O or        —COalkyl;    -   R₂ is hydrogen, alkyl or cycloalkyl;    -   R₃, at each occurrence, are independently hydrogen or alkyl;    -   R₄, at each occurrence, are independently hydrogen, halo,        hydroxy or alkyl;    -   R₅, at each occurrence, is hydrogen, halo, alkyl or cycloalkyl;    -   m, at each occurrence, is 0-2; and    -   n, at each occurrence, is 0-2.

In another embodiment, compounds of the present invention, or astereoisomer or pharmaceutically acceptable salt from thereof, are thosein which the compound is a compound of formula (Ia):

wherein

-   -   Ring A is an optionally substituted five to seven membered        heterocyclic ring;    -   T is

-   -   W is —OH;    -   R and R₁ are independently hydrogen, alkyl, halo, C═O or        —COalkyl;    -   R₂ is C₁-C₄alkyl or cycloalkyl;    -   R₃, at each occurrence, are independently hydrogen or alkyl;    -   R₄, at each occurrence, are independently hydroxy or C₁-C₄alkyl;    -   R₅, at each occurrence, is a halogen atom;    -   m, at each occurrence, is 0-2; and    -   n, at each occurrence, is 0-2.

In still yet another embodiment, compounds of the present invention, ora stereoisomer or pharmaceutically acceptable salt from thereof, arethose of compound Ib:

wherein

-   -   Ring A is an optionally substituted five to six membered        heterocyclic ring;    -   R and R₁ are independently hydrogen, alkyl, halo, C═O or        —COalkyl;    -   R₂ is C₁-C₄alkyl;    -   R₃, at each occurrence, are independently hydrogen or alkyl;    -   R₄, at each occurrence, are independently C₁-C₄alkyl;    -   R₅, at each occurrence, is —Cl;    -   m, at each occurrence, is 0-2; and    -   n, at each occurrence, is 0-2.

Representative compounds of the invention include the following:

-   (2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-8-oxa-6-azaspiro[4.5]decane-2-carboxamide,-   (2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6,8-diazaspiro[4.5]decane-2-carboxamide,-   N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6-oxa-8-azaspiro[4.5]decane-1-carboxamide,-   (7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxamide;-   (2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxamide-   (3R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3    ‘-methyl-6’-oxo-2′,5′,6′,7′-tetrahydro-spiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxamide,-   (7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxamide,-   (7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxamide,-   (7R)-3-chloro-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide,-   (7R)-1-acetyl-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide,-   N—((R)-1-((S)-4-(4-Chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-oxaspiro[2.4]heptane-5-carboxamide,    and-   N—((R)-1-((S)-4-(4-Chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-oxaspiro[3.4]octane-6-carboxamide.

In another embodiment, the present invention is directed to apharmaceutical composition, comprising a pharmaceutically acceptablecarrier and a therapeutically effective amount of a compound of thepresent invention.

In another embodiment, the present invention is directed to a method formodulation of chemokine or chemokine receptor activity comprisingadministering to a patient in need thereof a therapeutically effectiveamount of a compound of the present invention.

In another embodiment, the present invention is directed to a method formodulation of CCR-1 receptor activity comprising administering to apatient in need thereof a therapeutically effective amount of a compoundof the present invention.

In another embodiment, the present invention is directed to a method formodulation of MIP-1α, MCP-3, MCP-4, RANTES activity, preferablymodulation of MIP-1α activity, that is mediated by the CCR-1 receptorcomprising administering to a patient in need thereof a therapeuticallyeffective amount of a compound of the present invention.

In another embodiment, the present invention is directed to a method fortreating disorders, comprising administering to a patient in needthereof a therapeutically effective amount of a compound of the presentinvention, said wherein said disorder is selected from osteoarthritis,aneurysm, fever, cardiovascular effects, Crohn's disease, congestiveheart failure, autoimmune diseases, HIV-infection, HIV-associateddementia, psoriasis, idiopathic pulmonary fibrosis, transplantarteriosclerosis, physically- or chemically-induced brain trauma,inflammatory bowel disease, alveolitis, colitis, systemic lupuserythematosus, nephrotoxic serum nephritis, glomerularnephritis, asthma,multiple sclerosis, atherosclerosis, rheumatoid arthritis, restinosis,organ transplantation, psoriatic arthritis, multiple myeloma, allergies,for example, skin and mast cell degranulation in eye conjunctiva,hepatocellular carcinoma, osteoporosis, renal fibrosis and cancer,preferably, Crohn's disease, psoriasis, inflammatory bowel disease,systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis,multiple myeloma, allergies, for example, skin and mast celldegranulation in eye conjunctiva, hepatocellular carcinoma, osteoporosisand renal fibrosis.

In another embodiment, the present invention is directed to a method fortreating inflammatory diseases, for example, inflammatory diseases whichare at least partially mediated by CCR-1, comprising administering to apatient in need thereof a therapeutically effective amount of a compoundof the present invention.

In another embodiment, the present invention is directed to a method formodulation of CCR1 activity comprising administering to a patient inneed thereof a therapeutically effective amount of a compound of thepresent invention.

In another embodiment, the present invention is directed the use of acompound of the present invention in the preparation of a medicament forthe treatment of a disorder, said disorder is selected fromosteoarthritis, aneurysm, fever, cardiovascular effects, Crohn'sdisease, congestive heart failure, autoimmune diseases, HIV-infection,HIV-associated dementia, psoriasis, idiopathic pulmonary fibrosis,transplant arteriosclerosis, physically- or chemically-induced braintrauma, inflammatory bowel disease, alveolitis, colitis, systemic lupuserythematosus, nephrotoxic serum nephritis, glomerularnephritis, asthma,multiple sclerosis, atherosclerosis, rheumatoid arthritis, restinosis,organ transplantation, psoriatic arthritis, multiple myeloma, allergies,for example, skin and mast cell degranulation in eye conjunctiva,hepatocellular carcinoma, osteoporosis, renal fibrosis and cancer,preferably, Crohn's disease, psoriasis, inflammatory bowel disease,systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis,multiple myeloma, allergies, for example, skin and mast celldegranulation in eye conjunctiva, hepatocellular carcinoma, osteoporosisand renal fibrosis.

In another embodiment, the present invention is directed to a compoundof the present invention for use in therapy.

In another embodiment, the present invention is directed to apharmaceutical composition comprising a compound of the presentinvention and one or more active ingredients.

In another embodiment, the present invention is directed to a method formodulation of chemokine or chemokine receptor activity comprisingadministering to a patient in need thereof a therapeutically effectiveamount of a pharmaceutical composition comprised of a compound of thepresent invention and one or more active ingredients.

In another embodiment, the present invention is directed to a method formodulation of CCR-1 receptor activity comprising administering to apatient in need thereof a therapeutically effective amount of apharmaceutical composition comprised of a compound of the presentinvention and one or more active ingredients.

In yet another embodiment, the present invention is directed to a methodfor modulation of MIP-1α, MCP-3, MCP-4, RANTES activity, preferablymodulation of MIP-1α activity, that is mediated by the CCR-1 receptorcomprising administering to a patient in need thereof a therapeuticallyeffective amount of a pharmaceutical composition comprised of a compoundof the present invention and one or more active ingredients.

In another embodiment, the present invention is directed to a method fortreating a disorder, comprising administering to a patient in needthereof a therapeutically effective amount of a pharmaceuticalcomposition comprised of a compound of the present invention and one ormore active ingredients, wherein said disorder is selected fromosteoarthritis, aneurysm, fever, cardiovascular effects, Crohn'sdisease, congestive heart failure, autoimmune diseases, HIV-infection,HIV-associated dementia, psoriasis, idiopathic pulmonary fibrosis,transplant arteriosclerosis, physically- or chemically-induced braintrauma, inflammatory bowel disease, alveolitis, colitis, systemic lupuserythematosus, nephrotoxic serum nephritis, glomerularnephritis, asthma,multiple sclerosis, atherosclerosis, rheumatoid arthritis, restinosis,organ transplantation, psoriatic arthritis, multiple myeloma, allergies,for example, skin and mast cell degranulation in eye conjunctiva,hepatocellular carcinoma, osteoporosis, renal fibrosis and cancer,preferably, Crohn's disease, psoriasis, inflammatory bowel disease,systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis,multiple myeloma, allergies, for example, skin and mast celldegranulation in eye conjunctiva, hepatocellular carcinoma, osteoporosisand renal fibrosis.

In yet another embodiment, the present invention, is directed to amethod for treating inflammatory diseases, preferably, inflammatorydiseases which are at least partially mediated by CCR-1, comprisingadministering to a patient in need thereof a therapeutically effectiveamount of a pharmaceutical composition comprised of a compound ofpresent invention and one or more active ingredients.

In another embodiment, the present invention is directed to a method formodulation of CCR-1 activity comprising administering to a patient inneed thereof a therapeutically effective amount of a pharmaceuticalcomposition comprised of a compound of the present invention and one ormore active ingredients.

In another embodiment, the present invention is directed to the use of apharmaceutical composition comprised of a compound of the presentinvention and one or more active ingredients in the preparation of amedicament for the treatment of a disorder, said disorder is selectedfrom osteoarthritis, aneurysm, fever, cardiovascular effects, Crohn'sdisease, congestive heart failure, autoimmune diseases, HIV-infection,HIV-associated dementia, psoriasis, idiopathic pulmonary fibrosis,transplant arteriosclerosis, physically- or chemically-induced braintrauma, inflammatory bowel disease, alveolitis, colitis, systemic lupuserythematosus, nephrotoxic serum nephritis, glomerularnephritis, asthma,multiple sclerosis, atherosclerosis, rheumatoid arthritis, restinosis,organ transplantation, psoriatic arthritis, multiple myeloma, allergies,for example, skin and mast cell degranulation in eye conjunctiva,hepatocellular carcinoma, osteoporosis, renal fibrosis and cancer,preferably, Crohn's disease, psoriasis, inflammatory bowel disease,systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis,multiple myeloma, allergies, for example, skin and mast celldegranulation in eye conjunctiva, hepatocellular carcinoma, osteoporosisand renal fibrosis.

In still yet another embodiment, the present invention is directed tothe use of a pharmaceutical composition comprised of a compound of thepresent invention and one or more active ingredients in therapy.

In another embodiment, the present invention provides a compound of thepresent invention for use in therapy for treating an inflammatorydisease.

In another embodiment, the present invention provides a combinedpreparation of a compound of the present invention and additionaltherapeutic agent(s) for simultaneous, separate or sequential use intherapy.

In another embodiment, the present invention provides a combinedpreparation of a compound of the present invention and additionaltherapeutic agent(s) for simultaneous, separate or sequential use intreatment of an inflammatory disease.

In another embodiment, the present invention provides a novel article ofmanufacture, comprising: (a) a first container; (b) a pharmaceuticalcomposition located within the first container, wherein the composition,comprises: a first therapeutic agent, comprising: a compound of thepresent invention; and (c) a package insert stating that thepharmaceutical composition can be used for the treatment of aninflammatory disease.

In another preferred embodiment, the present invention provides a novelarticle of manufacture, further comprising: (d) a second container;wherein components (a) and (b) are located within the second containerand component (c) is located within or outside of the second container.

In another embodiment, the present invention provides a novel article ofmanufacture, comprising: (a) a first container; (b) a pharmaceuticalcomposition located within the first container, wherein the composition,comprises: a first therapeutic agent, comprising: a compound of thepresent invention; and (c) a package insert stating that thepharmaceutical composition can be used in combination with a secondtherapeutic agent to treat an inflammatory disease.

The invention may be embodied in other specific forms without departingfrom the spirit or essential attributes thereof. This invention alsoencompasses all combinations of alternative aspects of the inventionnoted herein. It is understood that any and all embodiments of thepresent invention may be taken in conjunction with any other embodimentto describe additional embodiments of the present invention.Furthermore, any elements of an embodiment may be combined with any andall other elements from any of the embodiments to describe additionalembodiments.

DEFINITIONS

The following are definitions of terms that may be used in thespecification. The initial definition provided for a group or termherein applies to that group or term throughout the specificationindividually or as part of another group, unless otherwise indicated.

The compounds herein described may have asymmetric centers. Compounds ofthe present invention containing an asymmetrically substituted atom maybe isolated in optically active or racemic forms. It is well known inthe art how to prepare optically active forms, such as by resolution ofracemic forms or by synthesis from optically active starting materials.Many geometric isomers of olefins, C═N double bonds, and the like canalso be present in the compounds described herein, and all such stableisomers are contemplated in the present invention. Cis and transgeometric isomers of the compounds of the present invention aredescribed and may be isolated as a mixture of isomers or as separatedisomeric forms. All chiral, diastereomeric, racemic forms and allgeometric isomeric forms of a structure are intended, unless thespecific stereochemistry or isomeric form is specifically indicated.

One enantiomer of a compound of Formula I may display superior activitycompared with the other. Thus, all of the stereochemistries areconsidered to be a part of the present invention. When required,separation of the racemic material can be achieved by HPLC using achiral column or by a resolution using a resolving agent such ascamphonic chloride as in Young, S. D. et al., Antimicrobial Agents andChemotherapy, 2602-2605 (1995).

The term “substituted,” as used herein, means that any one or morehydrogens on the designated atom or ring is replaced with a selectionfrom the indicated group, provided that the designated atom's or ringatom's normal valency is not exceeded, and that the substitution resultsin a stable compound. When a substituent is keto (i.e., ═O), then 2hydrogens on the atom are replaced.

When any variable (e.g., R₄) occurs more than one time in anyconstituent or formula for a compound, its definition at each occurrenceis independent of its definition at every other occurrence. Thus, forexample, if a group is shown to be substituted with (R₄)_(m) and m is0-3, then said group may optionally be substituted with up to three R₄groups and R₄ at each occurrence is selected independently from thedefinition of R₄. Also, combinations of substituents and/or variablesare permissible only if such combinations result in stable compounds.

When a bond to a substituent is shown to cross a bond connecting twoatoms in a ring, then such substituent may be bonded to any atom on thering. When a substituent is listed without indicating the atom via whichsuch substituent is bonded to the rest of the compound of a givenformula, then such substituent may be bonded via any atom in suchsubstituent. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds.

As used herein, “alkyl” is intended to include both branched andstraight-chain saturated aliphatic hydrocarbon groups containing 1 to 20carbons, preferably 1 to 10 carbons, more preferably 1 to 8 carbons, inthe normal chain, such as methyl, ethyl, propyl, isopropyl, butyl,t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl,octyl, 2,2,4-trimethyl-pentyl, nonyl, decyl, undecyl, dodecyl, thevarious branched chain isomers thereof, and the like as well as suchgroups may optionally include 1 to 4 substituents such as halo, forexample F, Br, Cl, or I, or CF₃, alkyl, alkoxy, aryl, aryloxy,aryl(aryl) or diaryl, arylalkyl, arylalkyloxy, alkenyl, cycloalkyl,cycloalkylalkyl, cycloalkylalkyloxy, amino, hydroxy, hydroxyalkyl, acyl,heteroaryl, heteroaryloxy, heteroarylalkyl, heteroarylalkoxy,aryloxyalkyl, alkylthio, arylalkylthio, aryloxyaryl, alkylamido,alkanoylamino, arylcarbonylamino, nitro, cyano, thiol, haloalkyl,trihaloalkyl, and/or alkylthio.

Unless otherwise indicated, the term “alkenyl” as used herein by itselfor as part of another group refers to straight or branched chainradicals of 2 to 20 carbons, preferably 2 to 12 carbons, and morepreferably 1 to 8 carbons in the normal chain, which include one to sixdouble bonds in the normal chain, such as vinyl, 2-propenyl, 3-butenyl,2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 2-heptenyl,3-heptenyl, 4-heptenyl, 3-octenyl, 3-nonenyl, 4-decenyl, 3-undecenyl,4-dodecenyl, 4,8,12-tetradecatrienyl, and the like, and which may beoptionally substituted with 1 to 4 substituents, namely, halogen,haloalkyl, alkyl, alkoxy, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl,amino, hydroxy, heteroaryl, cycloheteroalkyl, alkanoylamino, alkylamido,arylcarbonyl-amino, nitro, cyano, thiol, alkylthio, and/or any of thealkyl substituents set out herein.

Unless otherwise indicated, the term “alkynyl” as used herein by itselfor as part of another group refers to straight or branched chainradicals of 2 to 20 carbons, preferably 2 to 12 carbons and morepreferably 2 to 8 carbons in the normal chain, which include one triplebond in the normal chain, such as 2-propynyl, 3-butynyl, 2-butynyl,4-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl,4-heptynyl, 3-octynyl, 3-nonynyl, 4-decynyl, 3-undecynyl, 4-dodecynyl,and the like, and which may be optionally substituted with 1 to 4substituents, namely, halogen, haloalkyl, alkyl, alkoxy, alkenyl,alkynyl, aryl, arylalkyl, cycloalkyl, amino, heteroaryl,cycloheteroalkyl, hydroxy, alkanoylamino, alkylamido, arylcarbonylamino,nitro, cyano, thiol, and/or alkylthio, and/or any of the alkylsubstituents set out herein.

Unless otherwise indicated, the term “cycloalkyl” as employed hereinalone or as part of another group includes saturated or partiallyunsaturated (containing 1 or 2 double bonds) cyclic hydrocarbon groupscontaining 1 to 3 rings, including monocyclic alkyl, bicyclic alkyl (orbicycloalkyl) and tricyclic alkyl, containing a total of 3 to 20 carbonsforming the ring, preferably 3 to 10 carbons, forming the ring and whichmay be fused to 1 or 2 aromatic rings as described for aryl, whichincludes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, cyclodecyl and cyclododecyl, cyclohexenyl,

any of which groups may be optionally substituted with 1 to 4substituents such as halogen, alkyl, alkoxy, hydroxy, aryl, aryloxy,arylalkyl, cycloalkyl, alkylamido, alkanoylamino, oxo, acyl,arylcarbonylamino, amino, nitro, cyano, thiol, and/or alkylthio, and/orany of the substituents for alkyl.

Where alkyl groups as defined above have single bonds for attachment toother groups at two different carbon atoms, they are termed “alkylene”groups and may optionally be substituted as defined above for “alkyl”.

Where alkenyl groups as defined above and alkynyl groups as definedabove, respectively, have single bonds for attachment at two differentcarbon atoms, they are termed “alkenylene groups” and “alkynylenegroups”, respectively, and may optionally be substituted as definedabove for “alkenyl” and “alkynyl”.

“Halo” or “halogen” as used herein refers to fluoro, chloro, bromo, andiodo; and “haloalkyl” is intended to include both branched andstraight-chain saturated aliphatic hydrocarbon groups, for example CF₃,having the specified number of carbon atoms, substituted with 1 or morehalogen.

Unless otherwise indicated, the term “aryl” as employed herein alone oras part of another group refers to monocyclic and bicyclic aromaticgroups containing 6 to 10 carbons in the ring portion (such as phenyl ornaphthyl, including 1-naphthyl and 2-naphthyl) and may optionallyinclude 1 to 3 additional rings fused to a carbocyclic ring or aheterocyclic ring (such as aryl, cycloalkyl, heteroaryl, orcycloheteroalkyl rings

for example

and may be optionally substituted through available carbon atoms with 1,2, or 3 substituents, for example, hydrogen, halo, haloalkyl, alkyl,haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl,trifluoromethoxy, alkynyl, cycloalkyl-alkyl, cycloheteroalkyl,cycloheteroalkylalkyl, aryl, heteroaryl, arylalkyl, aryloxy,aryloxyalkyl, arylalkoxy, arylthio, arylazo, heteroarylalkyl,heteroarylalkenyl, heteroarylheteroaryl, heteroaryloxy, hydroxy, nitro,cyano, amino, substituted amino wherein the amino includes 1 or 2substituents (which are alkyl, aryl, or any of the other aryl compoundsmentioned in the definitions), thiol, alkylthio, arylthio,heteroarylthio, arylthioalkyl, alkoxyarylthio, alkylcarbonyl,arylcarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl,aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino,arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonylamino,or arylsulfonaminocarbonyl, and/or any of the alkyl substituents set outherein.

Unless otherwise indicated, the term “lower alkoxy”, “alkoxy”, “aryloxy”or “aralkoxy” as employed herein alone or as part of another groupincludes any of the above alkyl, aralkyl, or aryl groups linked to anoxygen atom.

Unless otherwise indicated, the term “amino” as employed herein alone oras part of another group refers to amino that may be substituted withone or two substituents, which may be the same or different, such asalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloheteroalkyl,cycloheteroalkylalkyl, cycloalkyl, cycloalkylalkyl, haloalkyl,hydroxyalkyl, alkoxyalkyl, or thioalkyl. These substituents may befurther substituted with a carboxylic acid and/or any of the R¹ groupsor substituents for R¹ as set out above. In addition, the aminosubstituents may be taken together with the nitrogen atom to which theyare attached to form 1-pyrrolidinyl, 1-piperidinyl, 1-azepinyl,4-morpholinyl, 4-thiamorpholinyl, 1-piperazinyl, 4-alkyl-1-piperazinyl,4-arylalkyl-1-piperazinyl, or 4-diarylalkyl-1-piperazinyl, all of whichmay be optionally substituted with alkyl, alkoxy, alkylthio, halo,trifluoromethyl, or hydroxy.

Unless otherwise indicated, the term “lower alkylthio,” “alkylthio,”“arylthio,” or “aralkylthio” as employed herein alone or as part ofanother group includes any of the above alkyl, aralkyl, or aryl groupslinked to a sulfur atom.

Unless otherwise indicated, the term “lower alkylamino,” “alkylamino,”“arylamino,” or “arylalkylamino” as employed herein alone or as part ofanother group includes any of the above alkyl, aryl, or arylalkyl groupslinked to a nitrogen atom.

As used herein, the term “heterocyclyl” or “heterocyclic system” isintended to mean a stable 5, 6, or 7-membered monocyclic or bicyclic or7, 8, 9, or 10-membered bicyclic heterocyclic ring which is saturated,partially unsaturated or unsaturated (aromatic), and which consists ofcarbon atoms and 1, 2, 3, or 4 heteroatoms independently selected fromthe group consisting of N, NH, O and S and including any bicyclic groupin which any of the above-defined heterocyclic rings is fused to abenzene ring. The nitrogen and sulfur heteroatoms may optionally beoxidized. The heterocyclic ring may be attached to its pendant group atany heteroatom or carbon atom, which results in a stable structure. Theheterocyclic rings described herein may be substituted on carbon or on anitrogen atom if the resulting compound is stable. If specificallynoted, a nitrogen in the heterocycle may optionally be quaternized. Itis preferred that when the total number of S and O atoms in theheterocycle exceeds 1, then these heteroatoms are not adjacent to oneanother. As used herein, the term “aromatic heterocyclic system” or“heteroaryl” is intended to mean a stable 5- to 7-membered monocyclic orbicyclic or 7- to 10-membered bicyclic heterocyclic aromatic ring whichconsists of carbon atoms and from 1 to 4 heteroatoms independentlyselected from the group consisting of N, O and S and is aromatic innature.

Examples of heterocycles include, but are not limited to, 1H-indazole,2-pyrrolidonyl, 2H,6H-1,5,2-dithiazinyl, 2H-pyrrolyl, 1H-indolyl,4-piperidonyl, 4aH-carbazole, 4H-quinolizinyl, 6H-1,2,5-thiadiazinyl,acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl,benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl,benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazalonyl,carbazolyl, 4aH-carbazolyl, β-carbolinyl, chromanyl, chromenyl,cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl,imidazolinyl, imidazolyl, indazolyl, indolenyl, indolinyl, indolizinyl,indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl,isoindolyl, isoquinolinyl, benzimidazolyl, isothiazolyl, isoxazolyl,morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinylperimidinyl,phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl,phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl,piperidinyl, pteridinyl, piperidonyl, 4-piperidonyl, pteridinyl,purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl,pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl,pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolyl, quinazolinyl,quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, carbolinyl,tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl,thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl,triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl,1,3,4-triazolyl, tetrazolyl, and xanthenyl. In another aspect of theinvention, the heterocycles include, but are not limited to, pyridinyl,thiophenyl, furanyl, indazolyl, benzothiazolyl, benzimidazolyl,benzothiaphenyl, benzofuranyl, benzoxazolyl, benzisoxazolyl, quinolinyl,isoquinolinyl, imidazolyl, indolyl, isoidolyl, piperidinyl, piperidonyl,4-piperidonyl, piperonyl, pyrrazolyl, 1,2,4-triazolyl, 1,2,3-triazolyl,tetrazolyl, thiazolyl, oxazolyl, pyrazinyl, and pyrimidinyl. Alsoincluded are fused ring and spiro compounds containing, for example, theabove heterocycles.

Examples of heteroaryls are 1H-indazole, 2H,6H-1,5,2-dithiazinyl,indolyl, 4aH-carbazole, 4H-quinolizinyl, 6H-1,2,5-thiadiazinyl,acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl,benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl,benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazalonyl,carbazolyl, 4aH-carbazolyl, β-carbolinyl, chromanyl, chromenyl,cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl,imidazolinyl, imidazolyl, indazolyl, indolenyl, indolinyl, indolizinyl,indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl,isoindolyl, isoquinolinyl (benzimidazolyl), isothiazolyl, isoxazolyl,morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinylperimidinyl,phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl,phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl,piperidinyl, pteridinyl, piperidonyl, 4-piperidonyl, pteridinyl,purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl,pyrazolotriazinyl, pyridazinyl, pyridooxazole, pyridoimidazole,pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl,pyrrolinyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl,quinoxalinyl, quinuclidinyl, carbolinyl, tetrahydrofuranyl,tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl,1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl,1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl,thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl,tetrazolyl, and xanthenyl. In another aspect of the invention, examplesof heteroaryls are indolyl, benzimidazolyl, benzofuranyl,benzothiofuranyl, benzoxazolyl, benzthiazolyl, benztriazolyl,benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazalonyl,cinnolinyl, furanyl, imidazolyl, indazolyl, indolyl, isoquinolinylisothiazolyl, isoxazolyl, oxazolyl, pyrazinyl, pyrazolyl,pyrazolotriazinyl, pyridazinyl, pyridyl, pyridinyl, pyrimidinyl,pyrrolyl, quinazolinyl, quinolinyl, thiazolyl, thienyl, and tetrazolyl.

The term “heterocyclylalkyl” or “heterocyclyl” as used herein alone oras part of another group refers to heterocyclyl groups as defined abovelinked through a C atom or heteroatom to an alkyl chain.

The term “heteroarylalkyl” or “heteroarylalkenyl” as used herein aloneor as part of another group refers to a heteroaryl group as definedabove linked through a C atom or heteroatom to an alkyl chain, alkylene,or alkenylene as defined above.

The term “cyano” as used herein, refers to a —CN group.

The term “nitro” as used herein, refers to an —NO₂ group.

The term “hydroxy” as used herein, refers to an OH group.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, “pharmaceutically acceptable salts” refer to derivativesof the disclosed compounds wherein the parent compound is modified bymaking acid or base salts thereof. Examples of pharmaceuticallyacceptable salts include, but are not limited to, mineral or organicacid salts of basic residues such as amines; alkali or organic salts ofacidic residues such as carboxylic acids; and the like. Thepharmaceutically acceptable salts include the conventional non-toxicsalts or the quaternary ammonium salts of the parent compound formed,for example, from non-toxic inorganic or organic acids. For example,such conventional non-toxic salts include those derived from inorganicacids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric,nitric and the like; and the salts prepared from organic acids such asacetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric,citric, ascorbic, palmidic, maleic, hydroxymaleic, phenylacetic,glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic,and the like.

The pharmaceutically acceptable salts of the present invention can besynthesized from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, nonaqueousmedia like ether, ethyl acetate, ethanol, isopropanol, or acetonitrileare preferred. Lists of suitable salts are found in Remington'sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa.,publ., p. 1418 (1985), the disclosure of which is hereby incorporated byreference.

The present invention is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include deuteriumand tritium. Isotopes of carbon include ¹³C and ¹⁴C.Isotopically-labeled compounds of the invention can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described herein, using an appropriateisotopically-labeled reagent in place of the non-labeled reagentotherwise employed.

Any compound that can be converted in vivo to provide the bioactiveagent (i.e., the compound of formula I) is a prodrug within the scopeand spirit of the invention.

The term “prodrugs” as employed herein includes esters, carbamates andcarbonates formed by reacting one or more hydroxyls of compounds offormula I with alkyl, alkoxy, or aryl substituted acylating agentsemploying procedures known to those skilled in the art to generateacetates, pivalates, methylcarbonates, benzoates, and the like.

Various forms of prodrugs are well known in the art and are describedin:

-   -   a) Wermuth, C. G. et al., The Practice of Medicinal Chemistry,        Ch. 31, Academic Press, publ. (1996);    -   b) Bundgaard, H., ed., Design of Prodrugs, Elsevier, publ.        (1985);    -   c) Krause, B. R. et al., Chapter 6: “ACAT Inhibitors:        Physiologic Mechanisms for Hypolipidemic and        Anti-Atherosclerotic Activities in Experimental Animals”,        Inflammation: Mediators and Pathways, CRC Press, Inc., publ.,        Ruffolo, Jr., R. R. et al., eds., pp. 173-198 (1995); and    -   d) Testa, B. et al., Hydrolysis in Drug and Prodrug Metabolism,        Wiley-VCH, publ. (2003).

These references are incorporated herein by reference.

In addition, compounds of the formula I are, subsequent to theirpreparation, preferably isolated and purified to obtain a compositioncontaining an amount by weight equal to or greater than 99% formula Icompound (“substantially pure” compound I), which is then used orformulated as described herein. Such “substantially pure” compounds ofthe formula I are also contemplated herein as part of the presentinvention.

All stereoisomers of the compounds of the instant invention arecontemplated, either in admixture or in pure or substantially pure form.The compounds of the present invention can have asymmetric centers atany of the carbon atoms including any one of the R substituents and/orexhibit polymorphism. Consequently, compounds of formula I can exist inenantiomeric, or diastereomeric forms, or in mixtures thereof. Theprocesses for preparation can utilize racemates, enantiomers, ordiastereomers as starting materials. When diastereomeric or enantiomericproducts are prepared, they can be separated by conventional methods forexample, chromatographic or fractional crystallization.

“Stable compound” and “stable structure” are meant to indicate acompound that is sufficiently robust to survive isolation to a usefuldegree of purity from a reaction mixture, and formulation into anefficacious therapeutic agent. The present invention is intended toembody stable compounds.

“Therapeutically effective amount” is intended to include an amount of acompound of the present invention alone or an amount of the combinationof compounds claimed or an amount of a compound of the present inventionin combination with other active ingredients effective to inhibit MIP-1αor effective to treat or prevent inflammatory disorders.

As used herein, “treating” or “treatment” cover the treatment of adisease-state in a mammal, particularly in a human, and include: (a)preventing the disease-state from occurring in a mammal, in particular,when such mammal is predisposed to the disease-state but has not yetbeen diagnosed as having it; (b) inhibiting the disease-state, i.e.,arresting it development; and/or (c) relieving the disease-state, i.e.,causing regression of the disease state.

Synthesis

The compounds of the present invention can be prepared in a number ofways well known to one skilled in the art of organic synthesis. Thecompounds of the present invention can be synthesized using the methodsdescribed below, together with synthetic methods known in the art ofsynthetic organic chemistry, or variations thereon as appreciated bythose skilled in the art. Preferred methods include, but are not limitedto, those described below. All references cited herein are herebyincorporated in their entirety herein by reference.

The novel compounds of this invention may be prepared using thereactions and techniques described in this section. The reactions areperformed in solvents appropriate to the reagents and materials employedand are suitable for the transformations being effected. Also, in thedescription of the synthetic methods described below, it is to beunderstood that all proposed reaction conditions, including choice ofsolvent, reaction atmosphere, reaction temperature, duration of theexperiment and work up procedures, are chosen to be the conditionsstandard for that reaction, which should be readily recognized by oneskilled in the art. It is understood by one skilled in the art oforganic synthesis that the functionality present on various portions ofthe molecule must be compatible with the reagents and reactionsproposed. Such restrictions to the substituents that are compatible withthe reaction conditions will be readily apparent to one skilled in theart and alternate methods must then be used. This will sometimes requirea judgment to modify the order of the synthetic steps or to select oneparticular process scheme over another in order to obtain a desiredcompound of the invention. It will also be recognized that another majorconsideration in the planning of any synthetic route in this field isthe judicious choice of the protecting group used for protection of thereactive functional groups present in the compounds described in thisinvention. An authoritative account describing the many alternatives tothe trained practitioner is Greene and Wuts (Protective Groups InOrganic Synthesis, Third Edition, Wiley and Sons, 1999).

Compounds of the type 1i of the present invention can be synthesized asshown in Scheme 1. Coupling of the keto acid 1a with amine 1b (WO2007092681) in the presence of a base such as triethylamine and acoupling reagent such as BOP or EDAC and HOBt affords the compounds ofthe type 1c. Keto acids of the type 1a are either commercially availableor can be made by methods well know in the art (see for example WO2007092681) Aminoallylation of ketone 1c (see for example, M. Sugiura,J. Am. Chem. Soc. 2004, 126, 7182) affords 1d. Protection of amine in 1dwith an appropriate protecting group (for e.g. tert-butoxycarbonyl orbenzyloxycarbonyl using Boc₂O or benzyloxycarbonyl chloride) followed byoxidative cleavage of the double bond using for example OsO₄/NaIO₄ orozone gas affords the aldehyde 1f. Reduction of the aldehyde 1f to thecorresponding alcohol 1g which can be accomplished using a reducingagent such as sodium borohydride, followed by de-protection of the aminoprotecting group (for example using an acid such as HCl for the Bocgroup) furnishes the amino alcohol 1h. Cyclization using a carbonylsource such as carbonyldiimidazole or dimethylcarbonate affords thecorresponding spirocyclic oxazolidinone 1i.

Compounds of the type 2h of the present invention can be synthesized asshown in Scheme 2. Aminoallylation of keto benzyl ester 2a (see forexample, M. Sugiura, J. Am. Chem. Soc. 2004, 126, 7182) providescompounds of type 2b. Keto benzyl ester of the type 2a can be made bymethods well know in the art (see for example WO 2007092681). Protectionof amine in 2b with an appropriate protecting group (for e.g.tert-butoxycarbonyl or benzyloxycarbonyl using Boc₂O orbenzyloxycarbonyl chloride) followed by oxidative cleavage of the doublebond using for example OsO₄/NaIO₄ or ozone gas affords the aldehyde 2d.Reductive amination of 2d (see for example, D. Dube, Tetrahedron Lett.1999, 40, 2295) followed by de-protection of the amino protecting group(for example using an acid such as HCl for the Boc group) furnishes thediamine 2e. Cyclization using a carbonyl source such ascarbonyldiimidazole or dimethylcarbonate affords the correspondingspirocyclic oxazolidinone 2f. Debenzylation of 2f under standardhydrogenation conditions yields the corresponding acid 2g. Coupling ofacid 2g with amine 1b (WO 2007092681) in the presence of a base such astriethylamine and a coupling reagent such as BOP or EDAC and HOBtaffords compounds of the type 2h.

Compounds of the type 3i of the present invention can be synthesized asshown in Scheme 3. Transesterification of keto ethyl ester 3a (see forexample, J. Matsuo, Tetrahedron: Asymmetry 2007, 18, 1906) yields thebenzyl ester 3b. Keto ethyl ester of type 3a are either commerciallyavailable or can be made by methods well known in the art. Allylation of3b (see for example, C. Li, Tetrahedron 1998, 54, 2347) followed byoxidative cleavage of the double bond using for example OsO₄/NaIO₄ orozone gas affords the aldehyde 3d. Reductive amination of 3d (see forexample, D. Dube, Tetrahedron Lett. 1999, 40, 2295) followed bydebenzylation under standard hydrogenation conditions affords acid 3f.Coupling of acid 3f with amine 1b (WO 2007092681) in the presence of abase such as triethylamine and a coupling reagent such as BOP or EDACand HOBt affords 3g. De-protection of the amino protecting group in 3gusing an acid such as HCl for the Boc group followed by cyclizationusing a carbonyl source such as carbonyldiimidazole or dimethylcarbonateaffords the corresponding spirocyclic oxazolidinone 3i.

Compounds of the type 4e of the present invention can be synthesized asshown in Scheme 4. Allylation of 2a yields the homoallyl alcohol 4a (seefor example, S. Silver, Eur. J. Org. Chem. 2005, 1058). Oxidation of thedouble bond in 4a affords acid 4b (see for example, R. Almquist, J. Med.Chem. 1985, 28, 1062). Curtis rearrangement of 4b affords theintermediate isocyanate which undergoes intramolecular cyclization toafford the oxazolidinone 4c. Debenzylation under standard hydrogenationconditions affords acid 4d. Coupling of acid 4d with amine 1b (WO2007092681) in the presence of a base such as triethylamine and acoupling reagent such as BOP or EDAC and HOBt affords the compounds ofthe type 4e.

Compounds of the type 5e of the present invention can be synthesized asshown in Scheme 5. Oxidative cleavage of the double bond in 4a using forexample ozone gas affords the aldehyde 5a. Reductive amination of 5a(see for example, D. Dube, Tetrahedron Lett. 1999, 40, 2295) affordsamine 5b. Cyclization of 5b using a carbonyl source such ascarbonyldiimidazole or dimethylcarbonate affords the correspondingspirocyclic oxazolidinone 5c, which upon debenzylation under standardhydrogenation conditions affords acid 5d. Coupling of acid 5d with amine1b (WO 2007092681) in the presence of a base such as triethylamine and acoupling reagent such as BOP or EDAC and HOBt affords the compounds ofthe type 5e.

Compounds of the type 6c of the present invention can be synthesized asshown in Scheme 6. Treatment of the keto benzylester 2a with anaminopyrazole using conditions described in (V. Lipson, Chemistry ofHeterocyclic Compounds, 2007, 43, 490) affords 6a. Debenzylation of 6aunder standard hydrogenation conditions yields acid 6b. Coupling of acid6b with amine 1b (WO 2007092681) in the presence of a base such astriethylamine and a coupling reagent such as BOP or EDAC and HOBtaffords the compounds of the type 6c.

Compounds of the type 7c of the present invention can be synthesized asshown in Scheme 7. Treatment of the ketobenzyl ester 2a using sodiumcyanide and ammonium carbonate (see for example, P. Kiviranta, Bioorg.Med. Chem. Lett. 2007, 17, 2448) affords the hydantoin 7a. Debenzylationof 7a under standard hydrogenation conditions yields acid 7b. Couplingof acid 7b with amine 1b (WO 2007092681) in the presence of a base suchas triethylamine and a coupling reagent such as BOP or EDAC and HOBtaffords the compounds of the type of 7c.

Compounds of the type 8c of the present invention can be synthesized asshown in Scheme 8. Treatment of the ketobenzyl ester 2a using a cyanidesource such as sodium or potassium cyanide and an amine such as methylamine under modified Strecker's conditions affords the intermediateaminonitrile, which upon cyclization and hydrolysis with potassiumcyanate affords the hydantoin 8a (see for example, G. Carrera, Journalof Heterocyclic Chemistry, 1992, 29, 847). Debenzylation of 8a understandard hydrogenation conditions yields acid 8b. Coupling of acid 8bwith amine 1b (WO 2007092681) in the presence of a base such astriethylamine and a coupling reagent such as BOP or EDAC and HOBtaffords the compounds of the type of 8c.

Compounds of the type 9d, 9e and 9f of the present invention can besynthesized as shown in Scheme 9. Phenyl selenium chloride inducedcyclization of 2b affords 9a. Debenzylation of 9a under standardhydrogenation conditions can either exclusively afford the acid 9c andor the chloro acid 9b. Coupling of acid 9b and 9c individually withamine 1b (WO 2007092681) in the presence of a base such as triethylamineand a coupling reagent such as BOP or EDAC and HOBt affords thecompounds of the type 9d and 9e. Acetylation of 9e under standardacetylation conditions yields compounds of the type 9f.

EXAMPLES

Abbreviations used in the Examples are defined as “2×” for twice, “3×”for thrice, “° C.” for degrees Celsius, “g” for gram or grams, “mmol”for millimolar, “mL” for milliliter or milliliters, “M” for molar, “min”for minute or minutes, “mg” for milligram or milligrams, “h” for hour orhours, “LC” for liquid chromatography, “HPLC” for high performanceliquid chromatography, “MS” for mass spectroscopy, “RT” for roomtemperature, “THF” for tetrahydrofuran, “Et₂O” for diethyl ether,“EtOAc” for ethyl acetate, “Na₂SO₄” for sodium sulfate, “CH₂Cl₂” formethylene chloride, “TFA” for trifluoroacetic acid, “sat.” forsaturated, “NaHCO₃” for sodium bicarbonate, “N” for normal, “NaOH” forsodium hydroxide, “MeOH” for methanol, “MgSO₄” for magnesium sulfate,“Hex” for hexane, “H₂O” for water, “HCl” for hydrochloric acid, “v/v”for volume to volume ratio. “D”, “L”, “R” and “S” are stereochemicaldesignations familiar to those skilled in the art. Chemical names werederived using ChemDraw Ultra, version 8.0.8. When this program failed toprovide a name for the exact structure in question, an appropriate namewas assigned using the same methodology utilized by the program.

Example 1(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-8-oxa-6-azaspiro[4.5]decane-2-carboxamide

Step A:(R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-oxocyclopentanecarboxamide

To a solution of(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,hydrogen chloride salt (WO 2007092681, 300 mg, 0.799 mmol) in THF (2 mL)was added (R)-3-oxocyclopentanecarboxylic acid (WO 2007092681, 102 mg,0.799 mmol), triethylamine (0.223 mL, 1.599 mmol) and BOP (354 mg, 0.799mmol) at room temperature. The resulting mixture was stirred at roomtemperature for 3 hours. The reaction mixture was concentrated andpartitioned between ethyl acetate (15 mL) and 1N hydrogen chloride (1mL). The organic layer was washed with water (3 mL), brine (2 mL), driedover anhydrous Na₂SO₄, concentrated under reduced pressure and theresidue was subjected to preparative HPLC(Shimadzu VP-ODS 20×100 mm, 8min. gradient, Solvent A: 10% MeOH, 90% H₂O, 0.1% TFA; Solvent B: 90%MeOH, 10% H₂O, 0.1% TFA; wavelength: 220 nM). The desired fractions wereconcentrated and freeze dried to yield(R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-oxocyclopentanecarboxamide(300 mg, 0.668 mmol, 84% yield) as a solid. MS (ESI⁺) m/z 449 M⁺

Step B:(1R)-3-allyl-3-amino-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)cyclopentanecarboxamide

To(R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-oxocyclopentanecarboxamide(from step A, 300 mg, 0.668 mmol) was added ammonia (954 μL, 6.68 mmol)followed by 2-allyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (168 mg, 1.0mmol) at room temperature. The resulting mixture was stirred at roomtemperature for 16 hours. The reaction mixture was concentrated underreduced pressure and the residue was subjected to preparativeHPLC(Shimadzu VP-ODS 20×100 mm, 8 min. gradient, Solvent A: 10% MeOH,90% H₂O, 0.1% TFA; Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA; wavelength:220 nM). The desired fractions were collected, concentrated andpartitioned between ethyl acetate (20 mL) and sat. aqueous sodiumbicarbonate (10 mL). The organic layer was dried over anhydrous Na₂SO₄and concentrated under reduced pressure to yield(1R)-3-allyl-3-amino-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)cyclopentanecarboxamide(150 mg, 0.30 mmol, 45.8% yield) as a solid. MS (ESI⁺) m/z 490 M⁺

Step C: tert-butyl(3R)-1-allyl-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)cyclopentylcarbamate

To a solution of(1R)-3-allyl-3-amino-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)cyclopentanecarboxamide(from step B, 60 mg, 0.122 mmol) in dichloromethane (2 mL) was addeddi-tert-butyl dicarbonate (32 mg, 0.147 mmol) and triethylamine (0.034mL, 0.245 mmol). The resulting mixture was stirred at room temperaturefor 16 hours. The reaction mixture was diluted with dichloromethane (10mL) and sequentially washed with 1N hydrogen chloride (1 mL), water (5mL), brine (5 ML), dried over Na₂SO₄ and concentrated to yieldtert-butyl(3R)-1-allyl-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)cyclopentylcarbamate(70 mg, 0.119 mmol, 97% yield) as a clear oil. MS (ESI⁺) m/z 590 M⁺

Step D: tert-butyl(3R)-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-(2-oxoethyl)cyclopentylcarbamate

Tert-butyl(3R)-1-allyl-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)cyclopentylcarbamate(from step C, 70 mg, 0.119 mmol) was dissolved in dichloromethane (5 mL)and ozonized at −78° C. until the blue color persists. The reactionmixture was purged with nitrogen gas until the blue color disappeared.To the reaction mixture was added triethylamine (0.5 mL) and thecontents allowed to warm up to room temperature and stirred at roomtemperature for an additional 1 hour. The reaction mixture was passedthrough a 4g silica gel cartridge (eluted with ethyl acetate) and theeluate was concentrated to yield t-butyl(3R)-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-(2-oxoethyl)cyclopentylcarbamate(60 mg, 0.101 mmol, 85% yield) as an oil. MS (ESI⁺) m/z 592 M⁺

Step E: tert-butyl(3R)-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-(2-hydroxyethyl)cyclopentylcarbamate

To a solution of tert-butyl(3R)-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-(2-oxoethyl)cyclopentylcarbamate(from step D, 60 mg, 0.101 mmol) in MeOH (2 mL) at −78° C. was addedNaBH₄ (7.67 mg, 0.203 mmol). The resulting mixture was stirred at −78°C. for 1 hour. The solvent was removed under reduced pressure and to theresidue was added ethyl acetate (10 mL). The resulting mixture waswashed with water (2 mL), brine (2 mL), dried over Na₂SO₄ andconcentrated to yield t-butyl(3R)-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-(2-hydroxyethyl)cyclopentylcarbamate(58 mg, 0.098 mmol, 96% yield) as a solid. MS (ESI⁺) m/z 594 M⁺

Step F:(1R)-3-amino-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-(2-hydroxyethyl)cyclopentanecarboxamide

To t-butyl(3R)-3-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-(2-hydroxyethyl)cyclopentylcarbamate(58 mg, 0.098 mmol) was added hydrogen chloride (4M in dioxane, 0.5 mL).The resulting mixture was stirred at room temperature for 1 hour. Thereaction mixture was concentrated to yield(1R)-3-amino-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-(2-hydroxyethyl)cyclopentanecarboxamide,hydrogen chloride salt (50 mg, 0.094 mmol, 97% yield) as an oil. It wasused as such for the next step without further purification. MS (ESI⁺)m/z 494 M⁺

Step G:(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-8-oxa-6-azaspiro[4.5]decane-2-carboxamide

To a solution of(1R)-3-amino-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-(2-hydroxyethyl)cyclopentanecarboxamide(from step F, 30 mg, 0.061 mmol) in THF (1 mL) was added carbonyldiimidazole (14.77 mg, 0.091 mmol) and triethylamine (0.021 mL, 0.152mmol). The resulting mixture was stirred at room temperature for 4hours. The reaction mixture was concentrated and subjected preparativeHPLC(Shimadzu VP-ODS 20×100 mm, 8 min. gradient, Solvent A: 10% MeOH,90% H₂O, 0.1% TFA; Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA; wavelength:220 nM) to yield the desired product as a diastereomer mixture. Thediastereomer mixture was subjected to chiral preparative HPLC (Column:ChiralPak ASH, 250×3 cm, 5 μm; Flow rate: 120 mL/min; Mobile Phase:CO₂/MeOH=80/20; Inj. Vol.=2.0 mL, Wavelength=220 nM; isomer 1, retentiontime=5.75 min; isomer 2, retention time=9.10 min) to yield theindividual diasteromers (2 mg for each isomer, 3.85 μmol, 6.33% yield)as a solid.

Isomer 1:

MS (ESI⁺)=520 M⁺; Purity: >95%; 1H NMR (500 MHz, CHLOROFORM-d)[rotamers, Integration values are assigned based on a 1:1 inseparablemixture of a and b rotamers for a total of 74 protons, excluding the OHproton] δ ppm 0.77 (s, 6H) 0.81-0.99 (m, 12H) 1.05 (d, J=6.87 Hz, 3H)1.41-2.24 (m, 27H) 2.53-2.72 (m, J=17.91, 13.52, 13.52, 4.81 Hz, 2H)2.76-2.95 (m, J=9.83, 9.59, 9.59, 4.54 Hz, 1H) 3.18 (td, J=13.06, 2.75Hz, 1H) 3.38 (dd, J=13.20, 1.37 Hz, 1H) 3.61 (d, J=13.47 Hz, 1H)3.64-3.71 (m, 1H) 3.94 (dd, J=14.16, 2.89 Hz, 1H) 4.11 (dd, J=13.20,1.65 Hz, 1H) 4.21-4.37 (m, 2H) 4.65 (dd, J=12.37, 4.12 Hz, 1H) 4.96 (td,J=9.14, 5.64 Hz, 2H) 6.47 (d, J=8.80 Hz, 1H) 6.55 (d, J=8.80 Hz, 1H)6.99 (s, 1H) 7.13 (s, 1H) 7.29-7.50 (m, 8H); RP-HPLC: Retention time=3.1min (YMC S5 ODS 4.6×50 mm; 4 min gradient, run time 8 min, solvent A:10% MeOH, 90% H₂O, 0.2% H₃PO₄; Solvent B: 90% MeOH, 10% H₂O, 0.2%H₃PO₄);

Isomer 2:

MS (ESI⁺)=520 M⁺; Purity: >95%; RP-HPLC: Retention time=3.1 min (YMC S5ODS 4.6×50 mm; 4 min gradient, run time 8 min, solvent A: 10% MeOH, 90%H₂O, 0.2% H₃PO₄; Solvent B: 90% MeOH, 10% H₂O, 0.2% H₃PO₄.

Example 2(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6,8-diazaspiro[4.5]decane-2-carboxamide

Step A: (1R)-benzyl 3-allyl-3-aminocyclopentanecarboxylate

To a solution of (R)-benzyl 3-oxocyclopentanecarboxylate (WO 2007092681,1 g, 4.58 mmol) in methanol (5 mL) was added2-allyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.155 g, 6.87 mmol) andammonia (7N solution in MeOH, 8 mL). The resulting mixture was stirredat room temperature for 16 hours. The reaction mixture was concentratedand the residue was subjected to preparative HPLC (Shimadzu VP-ODS20×100 mm, 8 min. gradient, Solvent A: 10% MeOH, 90% H₂O, 0.1% TFA;Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA; wavelength: 220 nM). The desiredfractions were collected, concentrated and partitioned between ethylacetate (20 mL) and saturated aqueous sodium bicarbonate solution (10mL). The organic layer was dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure to yield (1R)-benzyl3-allyl-3-aminocyclopentanecarboxylate (616 mg, 2.38 mmol, 52% yield).MS (ESI⁺) m/z 260 (M+H)⁺

Step B: (1R)-benzyl 3-allyl-3-(tert-butoxycarbonylamino)cyclopentanecarboxylate

To a solution of (1R)-benzyl 3-allyl-3-aminocyclopentanecarboxylate(from step A, 370 mg, 1.427 mmol) in dichloromethane (5 mL) was addedtriethylamine (0.398 mL, 2.85 mmol) and di-tert-butyl dicarbonate (374mg, 1.712 mmol). The resulting mixture was stirred at room temperaturefor 16 hours. To the reaction mixture was added dichloromethane (20 mL)and the resulting mixture was washed with 1N hydrogen chloride (5 mL),water (5 ml), brine (5 mL), dried over Na₂SO₄ and concentrated to yield(1R)-benzyl 3-allyl-3-(tert-butoxycarbonylamino)cyclopentanecarboxylate(480 mg, 1.335 mmol, 94% yield) as an oil. MS (ESI⁺) m/z 382 (M+Na)⁺

Step C: (1R)-benzyl 3-(tert-butoxycarbonylamino)-3-(2-oxoethyl)cyclopentanecarboxylate

(1R)-benzyl 3-allyl-3-(tert-butoxycarbonylamino)cyclopentanecarboxylate(from step B, 100 mg, 0.278 mmol) was dissolved in dichloromethane (3mL) and ozonized at −78° C. until the blue color persists. The reactionmixture was purged with nitrogen gas until the blue color disappeared.To the reaction mixture was added triethylamine (0.5 mL) and thecontents allowed to warm up to room temperature and stirred at roomtemperature for additional 1 hour. The reaction mixture was passedthrough a 4 g silica gel cartridge (eluted with ethyl acetate) and theeluate was concentrated to yield (1R)-benzyl3-(tert-butoxycarbonylamino)-3-(2-oxoethyl)cyclopentanecarboxylate (80mg, 0.221 mmol, 80% yield) as an oil. It was used as such for the nextstep without further purification.

Step D: (1R)-benzyl 3-amino-3-(2-aminoethyl)cyclopentanecarboxylate

To a solution of (1R)-benzyl3-(tert-butoxycarbonylamino)-3-(2-oxoethyl)cyclopentanecarboxylate (fromstep C, 80 mg, 0.221 mmol) in acetonitrile (2 mL) was added tert-butylcarbamate (78 mg, 0.664 mmol), triethylsilane (77 mg, 0.664 mmol) andtrifluoroacetic acid (1.705 μL, 0.022 mmol). The resulting mixture wasstirred at room temperature for 4 hours. The reaction mixture wasconcentrated under reduced pressure to yield an oil which was treatedwith hydrogen chloride (4M in dioxane, 1 mL) at room temperature. Afterthe reaction was stirred at room temperature for 1 hour, the reactionmixture was concentrated and subjected to preparative HPLC(ShimadzuVP-ODS 20×100 mm, 8 min. gradient, Solvent A: 10% MeOH, 90% H₂O, 0.1%TFA; Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA; wavelength: 220 nM). Thedesired fractions were concentrated under reduced pressure to yield(1R)-benzyl 3-amino-3-(2-aminoethyl)cyclopentanecarboxylate, 2trifluoroacetic acid salt (56.1 mg, 0.114 mmol, 51.7% yield in 2 steps)as an oil. MS (ESI⁺) m/z 263 (M+H)⁺

Step E: (2R)-benzyl 7-oxo-6,8-diazaspiro[4.5]decane-2-carboxylate

To a solution of (1R)-benzyl3-amino-3-(2-aminoethyl)cyclopentanecarboxylate (from step D, 30 mg,0.114 mmol) in toluene (2 mL) was added triethylamine (0.048 mL, 0.343mmol) and 1,1′-Carbonyldiimidazole (37.1 mg, 0.229 mmol). The resultingmixture was stirred at 80° C. for 2 hours. To the reaction mixture wasadded ethyl acetate (10 mL) and the resulting mixture was washed with 1Nhydrogen chloride (1 mL), water (2 mL), brine (2 mL), dried over Na₂SO₄and concentrated under reduced pressure to yield (2R)-benzyl7-oxo-6,8-diazaspiro[4.5]decane-2-carboxylate (20 mg, 0.069 mmol, 60.7%yield) as an oil. It was used as such for the subsequent step withoutfurther purification. MS (ESI⁺) m/z 288 M⁺

Step F: (2R)-7-oxo-6,8-diazaspiro[4.5]decane-2-carboxylic acid

To a solution of (2R)-benzyl7-oxo-6,8-diazaspiro[4.5]decane-2-carboxylate (from step E, 20 mg, 0.069mmol) in methanol (2 mL) was added palladium (Pd/C, 10% wt., 7.38 mg,6.94 mmol). The resulting mixture was stirred under a hydrogen balloonfor 30 min. The reaction mixture was filtered. The filtrate wasconcentrated under reduced pressure to yield(2R)-7-oxo-6,8-diazaspiro[4.5]decane-2-carboxylic acid (10 mg, 0.050mmol, 72.7% yield) as an oil. It was used as such for the next stepwithout further purification. MS m/z 199 (M+H)⁺

Step G:(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6,8-diazaspiro[4.5]decane-2-carboxamide

To a solution of (2R)-7-oxo-6,8-diazaspiro[4.5]decane-2-carboxylic acid(from step F, 10 mg, 0.050 mmol) in THF (1 mL) was added(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,hydrogen chloride salt (34.2 mg, 0.091 mmol), triethylamine (7.03 μL,0.050 mmol) and BOP (22.31 mg, 0.050 mmol). The resulting mixture wasstirred at room temperature for 2 hours. The reaction mixture wassubjected to preparative HPLC(Shimadzu VP-ODS 20×100 mm, 8 min.gradient, Solvent A: 10% MeOH, 90% H₂O, 0.1% TFA; Solvent B: 90% MeOH,10% H₂O, 0.1% TFA; wavelength: 220 nM). The desired fractions wereconcentrated under reduced pressure and freeze dried to yield(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6,8-diazaspiro[4.5]decane-2-carboxamide(4 mg, solid, 7.71 mmol, 15.27% yield) as a mixture of diasteromers.RP-HPLC: Retention time=3.19 min (YMC S5 ODS 4.6×50 mm; 4 min gradient,run time 8 min, solvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄; Solvent B: 90%MeOH, 10% H2O, 0.2% H₃PO₄); Purity: >95%; MS (ESI⁺) m/z 519 M⁺

Example 3N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6-oxa-8-azaspiro[4.5]decane-1-carboxamide

Step A: benzyl 2-oxocyclopentanecarboxylate

To ethyl 2-oxocyclopentanecarboxylate (4 g, 25.6 mmol) was added benzylalcohol (3.6 g, 33.3 mmol) and the resulting mixture was heated to 175°C. for 1 hour. The reaction mixture was concentrated under reducedpressure using a high vacuum pump to yield benzyl2-oxocyclopentanecarboxylate (5.2 g, 23.84 mmol, 93% yield) as an oil.This compound was used for the next step without further purification.

Step B: Benzyl 2-allyl-2-hydroxycyclopentanecarboxylate (homochiral)

To a solution of benzyl 2-oxocyclopentanecarboxylate (from step A, 5 g,22.91 mmol) in a mixture of methanol (20 ml) and water (60.00 ml) wasadded 3-bromoprop-1-ene (8.31 g, 68.7 mmol) and indium (1.081 ml, 68.7mmol). The resulting mixture was stirred at room temperature for 16hours. The reaction mixture was concentrated and to the residue wasadded 1N hydrogen chloride (15 mL) and stirred at room temperature for 5min. The reaction mixture was partitioned between ethyl acetate (30 mL)and water (4 mL). The ethyl acetate layer is washed with brine (4 mL),dried over anhydrous Na₂SO₄ and concentrated under the reduced pressure.The residue was subjected to preparative HPLC (Shimadzu VP-ODS 20×100mm, 8 min. gradient, Solvent A: 10% MeOH, 90% H₂O, 0.1% TFA; Solvent B:90% MeOH, 10% H₂O, 0.1% TFA; wavelength: 220 nM). The desired fractionswere collected, concentrated under reduced pressure to yield the desiredproduct as a diastereomer mixture. The diastereomer mixture wassubjected to chiral preparative HPLC to yield four individual isomers.The fractions corresponding to the second isomer was concentrated andfreeze dried to yield a solid (800 mg, 3.08 mmol, 13.4% yield). ChiralHPLC method: Column. ChiralPak AD-H, 250×3 cm, 5 μm; Flow rate: 130mL/min; Mobile Phase: CO₂/MeOH=70/30; Inj. Vol.=0.7 mL, Wavelength=220nM,: Retention time=3.18 min. MS (ESI⁺)=283 (M+Na)⁺

Step C: Benzyl 2-hydroxy-2-(2-oxoethyl)cyclopentanecarboxylate((homochiral)

Benzyl 2-allyl-2-hydroxycyclopentanecarboxylate (from step B, 150 mg,0.576 mmol) was dissolved in dichloromethane (5 mL) and ozonized at −78°C. until the blue color persists. The reaction mixture was purged withnitrogen gas until the blue color disappeared. To the reaction mixturewas added triethylamine (0.5 mL) and the contents allowed to warm up toroom temperature and stirred at room temperature for an additional 1hour. The reaction mixture was passed through a 4 g silica gel cartridge(eluted with ethyl acetate) and the eluate was concentrated to yieldbenzyl 2-hydroxy-2-(2-oxoethyl)cyclopentanecarboxylate (120 mg, 0.457mmol, 79% yield) as an oil. It was used as such for the next stepwithout further purification.

Step D: Benzyl2-(2-(tert-butoxycarbonylamino)ethyl)-2-hydroxycyclopentanecarboxylate(homochiral)

To a solution of benzyl 2-hydroxy-2-(2-oxoethyl)cyclopentanecarboxylate(from step C, 120 mg, 0.457 mmol) in acetonitrile (2 mL) was addedtert-butyl carbamate (214 mg, 1.830 mmol), triethylsilane (213 mg, 1.830mmol) and trifluoroacetic acid (0.070 mL, 0.915 mmol). The resultingmixture was stirred at room temperature for 3 hours. The reactionmixture was concentrated and subjected to preparative HPLC(ShimadzuVP-ODS 20×100 mm, 8 min. gradient, Solvent A: 10% MeOH, 90% H₂O, 0.1%TFA; Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA; wavelength: 220 nM). Thedesired fractions were concentrated and freeze dried to yield benzyl2-(2-(tert-butoxycarbonylamino)ethyl)-2-hydroxycyclopentanecarboxylate(70 mg, 0.193 mmol, 42.1% yield) as a solid. MS (ESI⁺)=386 (M+Na)⁺

Step E:2-(2-(t-butoxycarbonylamino)ethyl)-2-hydroxycyclopentanecarboxylic acid(homochiral)

To a solution of benzyl2-(2-(t-butoxycarbonylamino)ethyl)-2-hydroxycyclopentanecarboxylate(from step D, 70 mg, 0.193 mmol) in MeOH (3 mL) was added palladium (10%wt Pd/C, 20.50 mg, 0.019 mmol). The resulting mixture was stirred undera hydrogen balloon for 30 min. The reaction mixture was filtered. Thefiltrate was concentrated under reduced pressure to yield2-(2-(tert-butoxycarbonylamino)ethyl)-2-hydroxycyclopentanecarboxylicacid (50 mg, 0.183 mmol, 95% yield) as an oil. MS=273 (M+H)⁺

Step F: t-butyl2-(2-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-hydroxycyclopentyl)ethylcarbamate

To a solution of(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,hydrogen chloride salt (68.7 mg, 0.183 mmol) in THF (2 mL) was added2-(2-(t-butoxycarbonylamino)ethyl)-2-hydroxycyclopentanecarboxylic acid(from step E, 50 mg, 0.183 mmol), triethylamine (0.051 mL, 0.366 mmol)and BOP (81 mg, 0.183 mmol). The resulting mixture was stirred at roomtemperature for 16 hours. The solvent was removed under reducedpressure. To the residue was added ethyl acetate (20 mL) and theresulting mixture was washed with water (5 mL), brine (5 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to yieldtert-butyl2-(2-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-hydroxycyclopentyl)ethylcarbamateas an oil (100 mg, 0.168 mmol, 92% yield). It was used as such for thenext step without further purification. MS (ESI⁺)=494 (M-Boc)⁺

Step G:2-(2-aminoethyl)-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-hydroxycyclopentanecarboxamide

To t-butyl2-(2-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamoyl)-1-hydroxycyclopentyl)ethylcarbamate(from step F, 100 mg, 0.168 mmol) was added hydrogen chloride (4M indioxane, 0.5 mL). The resulting mixture was stirred at room temperaturefor 1 hour. The reaction mixture was concentrated and the residue waspartitioned between ethyl acetate (10 mL) and saturated aqueous sodiumbicarbonate solution (5 mL). The ethyl acetate layer was concentratedunder reduced pressure and the residue was subjected to preparative HPLC(Shimadzu VP-ODS 20×100 mm, 8 min. gradient, Solvent A: 10% MeOH, 90%H₂O, 0.1% TFA; Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA; wavelength: 220nM). The desired fractions were concentrated under reduced pressure andfreeze dried to yield2-(2-aminoethyl)-N-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-hydroxycyclopentanecarboxamide,trifluoroacetic acid salt (60 mg, 0.099 mmol, 58.6% yield) as a solid.MS (ESI⁺)=494 M⁺

Step H:N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6-oxa-8-azaspiro[4.5]decane-1-carboxamide

To a solution of2-(2-aminoethyl)-N-((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-hydroxycyclopentanecarboxamide(from step G, 30 mg, 0.061 mmol) in THF (0.5 mL) was added triethylamine(0.017 mL, 0.121 mmol) and 1,1′-Carbonyldiimidazole (14.77 mg, 0.091mmol) at room temperature. The resulting mixture was stirred at roomtemperature for 4 hours. The reaction mixture was subjected topreparative HPLC (Shimadzu VP-ODS 20×100 mm, 8 min. gradient, Solvent A:10% MeOH, 90% H2O, 0.1% TFA; Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA).The fractions corresponding to each individual diasteromer werecollected, concentrated and freeze dried to yield (isomer 1: 8 mg, 0.015mmol, 25% yield; isomer 2: 6 mg, 0.011 mmol, 19% yield).

Isomer 1:

Anal. RP-HPLC: Retention time=2.96 min (YMC S5 ODS 4.6×50 mm; gradientsolvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄; Solvent B: 90% MeOH, 10% H₂O,0.2% H₃PO₄). Purity: >95%; MS (ESI⁺)=520 M⁺;

Isomer 2:

Anal. RP-HPLC: Retention time=3.07 min (YMC S5 ODS 4.6×50 mm; gradientsolvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄; Solvent B: 90% MeOH, 10% H₂O,0.2% H₃PO₄); 1H NMR (400 MHz, CHLOROFORM-d) [rotamers, integrationvalues are assigned based on a 1:1 inseparable mixture of a and brotamers for a total of 74 protons, excluding the OH proton] δ ppm 0.76(s, 3H) 0.80 (s, 3H) 0.84 (s, 3H) 0.86 (s, 3H) 0.88-0.98 (m, 9H) 1.03(d, J=6.53 Hz, 3H) 1.45 (d, J=14.05 Hz, 1H) 1.60 (d, J=13.55 Hz, 1H)1.66-2.31 (m, 13H) 2.31-2.46 (m, 2H) 2.55-2.77 (m, 4H) 3.05-3.27 (m, 2H)3.42 (d, J=13.30 Hz, 4H) 3.55-3.80 (m, 2H) 3.93-4.15 (m, 4H) 4.26 (br.s., 2H) 4.64 (d, J=13.80 Hz, 2H) 4.92-5.05 (m, 3H) 6.60 (d, 1H)6.90-7.11 (m, 1H) 7.28-7.44 (m, 8H); MS (ESI⁺)=520 M⁺

Example 4(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxamide

Step A: (1R)-benzyl 3-allyl-3-hydroxycyclopentanecarboxylate

To a solution of (R)-benzyl 3-oxocyclopentanecarboxylate (1 g, 4.58mmol) in THF (10 mL) was added zinc powder (2.098 mL, 229 mmol),saturated aqueous ammonium chloride solution (20 mL) followed by3-bromoprop-1-ene (1.663 g, 13.75 mmol) slowly at room temperature. Theresulting mixture was heated to 80° C. for 3 hours, then allowed to stirat room temperature for additional 12 hours. The reaction mixture wasfiltered and the filtrate concentrated. To the residue was added ethylacetate (20 mL) and the resulting mixture was washed with water (5 ml),brine (5 mL), dried over anhydrous Na₂SO₄ and concentrated under reducedpressure to yield (1R)-benzyl 3-allyl-3-hydroxycyclopentanecarboxylate(580 mg, 2.228 mmol, 48.6% yield) as an oil. It was used as such for thenext step without further purification. MS (ESI⁺) m/z 283 (M+Na)⁺

Step B: 2-((3R)-3-(benzyloxycarbonyl)-1-hydroxycyclopentyl)acetic acid

To a solution of benzyl 3-allyl-3-hydroxycyclopentanecarboxylate (fromstep A, 150 mg, 0.576 mmol) in ethyl acetate (5 mL) was added ruthenium(IV) oxide hydrate (104 mg, 0.691 mmol) followed by 10% wt. aqueoussodium periodate solution (˜3 mL). The resulting mixture was stirred atroom temperature for 4 hours. Isopropyl alcohol (3 mL) was added toquench the reaction. The reaction mixture was filtered, and the filtratewas concentrated under reduced pressure to yield2-(3-(benzyloxycarbonyl)-1-hydroxycyclopentyl)acetic acid (130 mg, 0.467mmol, 81% yield) as an oil. It was used as such for the next stepwithout further purification. MS (ESI⁺) m/z 301 (M+Na)⁺

Step C: (7R)-benzyl 2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxylate

To a solution of2-((3R)-3-(benzyloxycarbonyl)-1-hydroxycyclopentyl)acetic acid (fromstep B, 130 mg, 0.467 mmol) in a mixture of toluene (2 mL) andtetrahydrofuran (1.0 mL) was added triethylamine (0.098 mL, 0.701 mmol)and diphenylphosphoryl azide (154 mg, 0.561 mmol) at room temperature.The resulting mixture was heated to 80° C. for 45 min. The reactionmixture was concentrated and the residue was subjected to preparativeHPLC (Shimadzu VP-ODS 20×100 mm, 8 min. gradient, Solvent A: 10% MeOH,90% H₂O, 0.1% TFA; Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA). Thefractions corresponding to the individual diasteromers were collected,concentrated, freeze dried and taken forward individually. Isomer 1: 20mg, 0.073 mmol, 15.5% yield, MS (ESI⁺) m/z 276 (M+H)⁺; Isomer 2: 20 mg,0.073 mmol, 15.5% yield, MS (ESI⁺) m/z 276 (M+H)⁺

Step D: (7R)-2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxylic acid

To a solution of (7R)-benzyl2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxylate (20 mg, 0.073 mmol) inmethanol (3 mL) was added palladium on carbon (10% wt., 15.46 mg, 0.015mmol). The resulting mixture was stirred under a hydrogen balloon for 30min. The reaction mixture was filtered and the filtrate was concentratedto yield (7R)-2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxylic acid (13 mgfor each isomer, 0.070 mmol, 97% yield) as an oil.

Step E:(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxamide

To a solution of(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,hydrogen chloride salt (26.3 mg, 0.070 mmol) in THF (1 mL) was added(7R)-2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxylic acid (13 mg, 0.070mmol), triethylamine (9.78 μL, 0.070 mmol) and BOP (31.0 mg, 0.070mmol). The resulting mixture was stirred at room temperature for 4hours. The reaction mixture was concentrated and the residue waspartitioned between ethyl acetate (10 mL) and saturated sodiumbicarbonate solution (4 mL). The ethyl acetate layer was concentratedunder reduced pressure and the residue was subjected to preparative HPLC(Shimadzu VP-ODS 20×100 mm, 8 min. gradient, Solvent A: 10% MeOH, 90%H2O, 0.1% TFA; Solvent B: 90% MeOH, 10% H2O, 0.1% TFA). The desiredfractions were collected, concentrated and freeze dried to yield(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxamide(15 mg for each isomer, 0.030 mmol, 42.2% yield) as a solid.

Isomer 1:

Anal. RP-HPLC: Retention time=3.12 min (YMC S5 ODS 4.6×50 mm; gradientsolvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄; Solvent B: 90% MeOH, 10% H₂O,0.2% H₃PO₄); Purity: >95%; MS (ESI⁺)=506 M;

Isomer 2:

Anal. RP-HPLC: Retention time=3.16 min (YMC S5 ODS 4.6×50 mm; gradientsolvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄; Solvent B: 90% MeOH, 10% H₂O,0.2% H₃PO₄); Purity: >95%; 1H NMR (400 MHz, CHLOROFORM-d) [rotamers,integration values are assigned based on a 1:1 inseparable mixture of aand b rotamers for a total of 70 protons, excluding the OH proton] δ ppm0.76 (d, J=2.01 Hz, 6H) 0.87 (d, J=4.27 Hz, 6H) 0.95 (td, J=12.42, 6.78Hz, 9H) 1.05 (d, J=6.53 Hz, 3H) 1.49 (d, J=14.31 Hz, 1H) 1.60 (d,J=14.05 Hz, 1H) 1.84-2.37 (m, 12H) 2.56-2.77 (m, J=13.99, 13.83, 13.83,4.77 Hz, 2H) 3.11 (d, J=12.80 Hz, 1H) 3.14-3.28 (m, 1H) 3.44 (d, J=12.80Hz, 1H) 3.53-3.79 (m, 5H) 4.02 (d, J=11.04 Hz, 1H) 4.11 (d, J=12.80 Hz,1H) 4.56-5.07 (m, 10H) 6.13-6.36 (m, 1H) 7.19 (d, J=9.03 Hz, 1H)7.28-7.49 (m, 8H); MS (ESI⁺)=506 M⁺

Example 5(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxamide

Step A: (1R)-benzyl 3-hydroxy-3-(2-oxoethyl)cyclopentanecarboxylate

(1R)-benzyl 3-allyl-3-hydroxycyclopentanecarboxylate (from step A,example 04, 200 mg, 0.768 mmol) was dissolved in dichloromethane (5 mL)and ozonized at −78° C. until the blue color persists. The reactionmixture was purged with nitrogen gas until the blue color disappeared.To the reaction mixture was added triethylamine (0.5 mL) and thecontents allowed to warm up to room temperature and stirred at roomtemperature for additional 1 hour. The reaction mixture was passedthrough a 4 g silica gel cartridge (eluted with ethyl acetate) and theeluant was concentrated under reduced pressure to yield (1R)-benzyl3-hydroxy-3-(2-oxoethyl)cyclopentanecarboxylate (100 mg, 0.381 mmol,49.6% yield) as an oil. It was used as such for the next step withoutfurther purification.

Step B: (1R)-benzyl 3-(2-aminoethyl)-3-hydroxycyclopentanecarboxylate

To a solution of (1R)-benzyl3-hydroxy-3-(2-oxoethyl)cyclopentanecarboxylate (from step A, 100 mg,0.381 mmol) in acetonitrile (2 mL) was added tert-butyl carbamate (134mg, 1.144 mmol), triethylsilane (133 mg, 1.144 mmol) and trifluoroaceticacid (2.94 μL, 0.038 mmol). The resulting mixture was stirred at roomtemperature for 4 hours. The reaction mixture was concentrated andtreated with hydrogen chloride (4M solution in dioxane, 2 mL)subsequently. After the reaction was stirred at room temperature for 30min, the reaction mixture was concentrated and subjected to preparativeHPLC (Shimadzu VP-ODS 20×100 mm, 8 min. gradient, Solvent A: 10% MeOH,90% H2O, 0.1% TFA; Solvent B: 90% MeOH, 10% H2O, 0.1% TFA). The desiredfractions were collected, concentrated and freeze dried to yield(1R)-benzyl 3-(2-aminoethyl)-3-hydroxycyclopentanecarboxylate,trifluoroacetic acid salt (40 mg, 0.106 mmol, 27.8% yield) as a solid.MS (ESI⁺)=264 M⁺

Step C: (2R)-benzyl 7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxylate

To a solution of (1R)-benzyl3-(2-aminoethyl)-3-hydroxycyclopentanecarboxylate, trifluoroacetic acidsalt (from step B, 35 mg, 0.093 mmol) in THF (0.5 mL) was addedtriethylamine (0.026 mL, 0.186 mmol) followed by1,1′-carbonyldiimidazole (22.56 mg, 0.139 mmol). The resulting mixturewas stirred at room temperature for 2 hours. The reaction mixture wasconcentrated and subjected to preparative HPLC (Shimadzu VP-ODS 20×100mm, 8 min. gradient, Solvent A: 10% MeOH, 90% H₂O, 0.1% TFA; Solvent B:90% MeOH, 10% H₂O, 0.1% TFA). The desired fractions were collected,concentrated and freeze dried to yield (2R)-benzyl7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxylate (22 mg, 0.076 mmol, 82%yield) as a solid. MS (ESI)=312 (M+Na)⁺

Step D: (2R)-7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxylic acid

To a solution of (2R)-benzyl7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxylate (from step C, 22 mg,0.076 mmol) in MeOH (1 mL) was added palladium on carbon (10% wt., 8.09mg, 7.60 μmol). The resulting mixture was stirred under a hydrogenballoon for 30 min. The reaction mixture was filtered and the filtrateconcentrated under reduced pressure to yield(2R)-7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxylic acid (15 mg, 0.075mmol, 99% yield) as an oil. MS

=200 (M+H)⁺

Step E:(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxamide

To a solution of (2R)-7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxylicacid (from step D, 18 mg, 0.090 mmol) in THF (0.5 mL) was added(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,hydrogen chloride salt (33.9 mg, 0.090 mmol), triethylamine (0.025 mL,0.181 mmol) and BOP (40.0 mg, 0.090 mmol). The resulting mixture wasstirred at room temperature for 16 hours. The reaction mixture wasconcentrated and subjected to preparative HPLC (Shimadzu VP-ODS 20×100mm, 8 min. gradient, Solvent A: 10% MeOH, 90% H2O, 0.1% TFA; Solvent B:90% MeOH, 10% H2O, 0.1% TFA). The desired fractions were collected,concentrated and freeze dried to yield the desired product as adiastereomer mixture. The diastereomer mixture was subjected to chiralpreparative HPLC (Column: ChiralPak AD-H, 250×3 cm, 5 μm; Flow rate: 110mL/min; Mobile Phase: CO2/IPA=65/35; Inj. Vol.=2.0 mL, Wavelength=220nM; isomer 1, retention time=4.04 min; isomer 2, retention time=13.80min) to yield the individual isomers (isomer 1: 20 mg, 0.038 mmol, 33.3%yield; isomer 2: 18 mg, 0.035 mmol, 30.0% yield) as a solid.

Isomer 1:

Anal. RP-HPLC: Retention time=3.07 min (YMC S5 ODS 4.6×50 mm; 4 mingradient, run time 8 min, solvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄;Solvent B: 90% MeOH, 10% H2O, 0.2% H₃PO₄). Purity: >95%; MS (ESI⁺) m/z520 M⁺;

Isomer 2:

Anal. RP-HPLC: Retention time=3.14 min (YMC S5 ODS 4.6×50 mm; 4 mingradient, run time 8 min, solvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄;Solvent B: 90% MeOH, 10% H2O, 0.2% H₃PO₄). Purity: >95%; MS (ESI⁺) m/z520 M⁺

Example 6(3R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3′-methyl-6′-oxo-2′,5′,6′,7′-tetrahydro-spiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxamide

Step A: (3R)-benzyl3′-methyl-6′-oxo-2′,5′,6′,7′-tetrahydrospiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxylate

To (R)-benzyl 3-oxocyclopentanecarboxylate (0.4 g, 1.833 mmol) weresequentially added N-methylpyrrolidone (3 mL),2,2-dimethyl-1,3-dioxane-4,6-dione (0.264 g, 1.833 mmol) and3-methyl-1H-pyrazol-5-amine (0.178 g, 1.833 mmol) at room temperature.The contents heated to 80° C. (oil bath temp.) for 75 min, and stirredat room temperature for additional 48 hours. The reaction mixture waspartitioned between ethyl acetate (40 mL) and 1N hydrogen chloride (15mL). The ethyl acetate layer was separated and sequentially washed with1N sodium hydroxide (15 mL), brine (20 mL), dried over sodium sulfate,concentrated and subjected to preparative HPLC (Phenomex Luna AXIA30×100 mm; 10 min. gradient; solvent A=10% MeOH, 90% H₂O, 0.1% TFA,solvent B=90% MeOH, 10% H₂O, 0.1% TFA). Fractions corresponding to thedesired product were isolated and concentrated. The residue that wasobtained was partitioned between dichloromethane (20 mL) and saturatedaqueous sodium bicarbonate solution. The dichloromethane layer wasseparated, dried over sodium sulfate and concentrated to yield a solid(33 mg, 0.097 mmol, 5.31%). It was used as such for the subsequent stepwithout further purification. MS (ESI) m/z 340.16 (M+H)⁺

Step B:(3R)-3′-methyl-6′-oxo-2′,5′,6′,7′-tetrahydrospiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxylicacid

To (3R)-benzyl3′-methyl-6′-oxo-2′,5′,6′,7′-tetrahydrospiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxylate(from step A: 0.033 g, 0.097 mmol) in ethyl acetate (5 mL) was addedPd—C (10% wt., 0.020 g, 0.019 mmol) under a nitrogen atmosphere. Thecontents were hydrogenated at room temperature using a hydrogen balloonfor 1.5 hours. The reaction mixture was concentrated; methanol (5 mL)was added followed by additional Pd—C (0.020 g, 0.019 mmol). Thecontents were hydrogenated at 25 psi for 1 hour. The reaction mixturewas filtered over a Whatman filter (FN MB 25 MM 0.45 uM) paper andwashed with methanol (2×10 mL). The filtrate was concentrated underreduced pressure to yield an oil (0.017 g) which was azoetroped with THF(2×10 mL) and used as such for the subsequent step without furtherpurification. MS (ESI) m/z 250.19 (M+H)⁺

Step C:(3R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3′-methyl-6′-oxo-2′,5′,6′,7′-tetrahydrospiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxamide

To(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,HCl (0.023 g, 0.060 mmol) in DMF (2 mL) were sequentially added(3R)-3′-methyl-6′-oxo-2′,5′,6′,7′-tetrahydrospiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxylicacid (from step B, 0.015 g, 0.060 mmol), N,N-diisopropylethylamine(0.021 mL, 0.120 mmol) and BOP (0.027 g, 0.060 mmol) at roomtemperature. The reaction mixture was stirred at room temperature for 18hours.

The reaction mixture was concentrated under reduced pressure (highvacuum pump) and subjected to preparative HPLC (Phenomex Luna AXIA30×100 mm; 10 min. gradient; solvent A=10% MeOH, 90% H₂O, 0.1% TFA,solvent B=90% MeOH, 10% H₂O, 0.1% TFA). Fractions corresponding to thedesired product were isolated and concentrated. The residue that wasobtained was partitioned between dichloromethane (20 mL) and saturatedaqueous sodium bicarbonate solution (10 mL). The dichloromethane layerwas separated, dried over sodium sulfate and concentrated to yield(3R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3′-methyl-6′-oxo-2′,5′,6′,7′-tetrahydrospiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxamide(0.008 g, 0.014 mmol, 23.32% yield) as a diastereomeric mixture. Anal.RP-HPLC: Retention time=3.24 min (YMC S5 Combi ODS 4.6×50 mm; 4 min.gradient; solvent A=10% MeOH, 90% H₂O, 0.2% H₃PO₄, solvent B=90% MeOH,10% H₂O, 0.2% H₃PO₄). Purity: >95%. MS (ESI) m/z 570.31 (M+H)+

Example 7(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxamide

Step A: (7R)-benzyl 2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylate

To (R)-benzyl 3-oxocyclopentanecarboxylate (0.5 g, 2.291 mmol) inethanol (5 mL) were sequentially added sodium cyanide (0.281 g, 5.73mmol), ammonium carbonate (2.86 g, 29.8 mmol) and water (5.00 mL) atroom temperature. The contents were heated at 70° C. (oil bath temp.)for 3 hours. The reaction mixture was kept at room temperature for 10min. Solid separates out. The solid was filtered and washed with water(3×10 mL). The solid was dried overnight under vacuum (155 mg, 0.538mmol, 23.5%) and used as such for the subsequent step without furtherpurification. MS (ESI) m/z 289.16 (M+H)⁺

Step B: (7R)-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylic acid

To (7R)-benzyl 2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylate (fromstep A, 0.14 g, 0.486 mmol) in methanol (10 mL) was added Pd—C (0.030 g,0.282 mmol) under a nitrogen atmosphere. The contents were hydrogenatedat 10 psi for 3 hours. The reaction mixture was filtered over a whatmanfilter (FN MB 25 MM 0.45 uM) paper and washed with methanol (2×5 mL).The filtrate was concentrated under reduced pressure to yield a solid(0.09 g, 0.454 mmol, 94% yield). It was used as such for the subsequentstep without further purification. MS m/z 197.3 (M−H)⁺

Step C:(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxamide

To(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,HCl (from step B, 0.085 g, 0.227 mmol) in THF (2 mL) was sequentiallyadded (7R)-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylic acid (stepB, 0.045 g, 0.227 mmol), N,N-diisopropylethylamine (0.079 mL, 0.454mmol) and BOP (0.100 g, 0.227 mmol) at room temperature. The contentswere stirred at room temperature for 20 hours. The reaction mixture wassubjected to preparative HPLC (Phenomex Luna AXIA 30×100 mm; 10 min.gradient; solvent A=10% MeOH, 90% H2O, 0.1% TFA, solvent B=90% MeOH, 10%H2O, 0.1% TFA) and fractions corresponding to desired product wereisolated and concentrated. The residue was partitioned betweendichloromethane (20 mL) and saturated aqueous sodium bicarbonatesolution (3 mL). The dichloromethane layer was dried over anhydroussodium sulfate and concentrated to yield a solid (0.095 g, 0.183 mmol,81% yield). Anal. RP-HPLC: Retention time=3.15 min (YMC S5 Combi ODS4.6×50 mm; 4 min. gradient; solvent A=10% MeOH, 90% H₂O, 0.2% H₃PO₄,solvent B=90% MeOH, 10% H2O, 0.2% H₃PO₄); Purity: >95%; mixture ofrotamers; MS (ESI) m/z 519 (M+H)⁺

Example 8(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxamide

Step A: (7R)-benzyl1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylate

To (R)-benzyl 3-oxocyclopentanecarboxylate (0.5 g, 2.291 mmol) weresequentially added ethanol (1.0 mL), water (0.5 mL) and methylaminehydrochloride (0.155 g, 2.291 mmol). The contents were stirred at roomtemperature for 20 min. Then potassium cyanide (0.149 g, 2.291 mmol) wasadded at room temperature and the contents stirred at room temperaturefor 60 hours. The reaction mixture was added to 2.3 mL of 1.0N hydrogenchloride taken in a 25 mL round bottom flask at room temperaturefollowed by the addition of 0.23 mL of concentrated hydrogen chloride atroom temperature. The reaction mixture was heated to 90° C. for 1 hour.The reaction mixture was concentrated under reduced pressure andpartitioned between water (5 mL) and ethyl acetate (15 mL). The ethylacetate layer was concentrated and subjected to preparative HPLC(Phenomex Luna AXIA 30×100 mm; 10 min. gradient; solvent A=10% MeOH, 90%H₂O, 0.1% TFA, solvent B=90% MeOH, 10% H₂O, 0.1% TFA). Fractionscorresponding to the desired product were concentrated, partitionedbetween ethyl acetate (15 mL) and saturated aqueous sodium bicarbonatesolution (10 mL). The ethyl acetate layer was separated, dried oversodium sulfate and concentrated to yield (7R)-benzyl1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylate (0.105 g,0.347 mmol, 15.16% yield) as an oil. It was used as such for thesubsequent step. MS (ESI) m/z 303.14 (M+H)⁺

Step B: (7R)-1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylicacid

To (7R)-benzyl1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylate (from step A,0.1 g, 0.331 mmol) in ethyl acetate (10 mL) was added Pd—C (0.035 g,0.033 mmol) under a nitrogen atmosphere. The contents were hydrogenatedat 20 psi for 1 hour. The reaction mixture was filtered over a Whatmanfilter (FN MB 25 MM 0.45 uM) paper and washed with ethyl acetate (2×5mL). The filtrate was concentrated under reduced pressure to yield anoil (0.05 g, 0.236 mmol, 71.2%) which was used in the next step withoutfurther purification. MS m/z 211.3 (M−H)⁺

Step C:(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxamide

To(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,HCl (0.088 g, 0.236 mmol) in THF (2 mL) were sequentially added(7R)-1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxylic acid(from step B, 0.05 g, 0.236 mmol), N,N-diisopropylethylamine (0.082 mL,0.471 mmol) and BOP (0.104 g, 0.236 mmol) at room temperature. Thecontents were stirred at room temperature for 20 hours. The reactionmixture was subjected to preparative HPLC (Phenomex Luna AXIA 30×100 mm;10 min. gradient; solvent A=10% MeOH, 90% H2O, 0.1% TFA, solvent B=90%MeOH, 10% H2O, 0.1% TFA) and fractions corresponding to the desiredproduct were isolated and concentrated. The residue was partitionedbetween dichloromethane (20 mL) and saturated aqueous sodium bicarbonatesolution (3 mL). The dichloromethane layer was dried over sodium sulfateand concentrated to yield the desired product as a diastereomer mixture.The diastereomer mixture was subjected to chiral preparative HPLC(Column: ChiralPak OD-H, 3×25 cm; Flow rate: 70 mL/min; Mobile Phase:CO2/MeOH=80/20; Wavelength=220 nM; isomer 1, retention time=14.62 min;isomer 2, retention time=21.05 min) to yield the individual isomers(isomer 1: 7 mg, 0.013 mmol, 15.5% yield; isomer 2: 11 mg, 0.021 mmol,24.4% yield) as a solid.

Isomer 1:

Anal. RP-HPLC: Retention time=3.20 min (YMC S5 ODS 4.6×50 mm; 4 mingradient, run time 8 min, solvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄;Solvent B: 90% MeOH, 10% H2O, 0.2% H₃PO₄). Purity: >95%; MS (ESI⁺) m/z533.21 M⁺;

Isomer 2:

Anal. RP-HPLC: Retention time=3.15 min (YMC S5 ODS 4.6×50 mm; 4 mingradient, run time 8 min, solvent A: 10% MeOH, 90% H₂O, 0.2% H₃PO₄;Solvent B: 90% MeOH, 10% H2O, 0.2% H₃PO₄). Purity: >95%; MS (ESI⁺) m/z533.23 M⁺

Example 9(7R)-3-chloro-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide

(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-4-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide

Step A: (7R)-benzyl 3-chloro-1-azaspiro[4.4]nonane-7-carboxylate

To phenylselenyl chloride (0.199 g, 1.041 mmol) was added (1R)-benzyl3-allyl-3-aminocyclopentanecarboxylate (example 2, step A, 0.090 g,0.347 mmol) dissolved in acetonitrile (2 mL). The contents were heatedat 80° C. for 18 hours. The reaction mixture was concentrated andpartitioned between ethyl acetate (20 mL) and saturated aqueous sodiumbicarbonate (10 mL). The ethyl acetate layer was dried over sodiumsulfate and concentrated. To the residue was added methanol (5 mL). Thesolid that separates was filtered and washed with methanol (2 mL). Tothe filtrate was added 4.0N hydrogen chloride in dioxane (1 mL) and thecontents concentrated. Ethyl acetate was added and the resulting mixturewas concentrated under reduced pressure. Methanol (5 mL) was added and asolid separates out The liquid was carefully pipetted out into a HPLCvial and subjected to preparative HPLC (Phenomex Luna AXIA 30×100 mm; 10min. gradient; solvent A=10% MeOH, 90% H₂O, 0.1% TFA, solvent B=90%MeOH, 10% H₂O, 0.1% TFA). Fractions corresponding to the desired productwere isolated and concentrated. The residue was partitioned betweendichloromethane (20 mL) and saturated aqueous sodium bicarbonatesolution (3 mL). The dichloromethane layer was dried over sodium sulfateand concentrated to yield (7R)-benzyl3-chloro-1-azaspiro[4.4]nonane-7-carboxylate (0.051 g, 0.174 mmol, 50.0%yield) as an oil. It was used as such for the subsequent step. MS (ESI)m/z 294.14 (M+H)⁺

Step B: (7R)-1-azaspiro[4.4]nonane-7-carboxylic acid (B-1) and(7R)-3-chloro-1-azaspiro[4.4]nonane-7-carboxylic acid (B-2)

To (7R)-benzyl 3-chloro-1-azaspiro[4.4]nonane-7-carboxylate (from stepA, 0.051 g, 0.174 mmol) in methanol (8 mL) was added Pd—C (0.020 g,0.019 mmol) (10%) under a nitrogen atmosphere. The contents werehydrogenated at 40 psi at room temperature for 3.5 hours. The reactionmixture was filtered over a Whatman filter (FN MB 25 MM 0.45 uM) paperand washed with methanol (2×5 mL). The filtrate was concentrated underreduced pressure to yield a mixture of B-1 and B-2 (0.039 g). It wasused as such for the subsequent step without further purification.

Step C:(7R)-3-chloro-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide(peak 1 and peak 2) and(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide(peak 3).

To a mixture of B-1 and B-2 (from step B, 0.039 g, 0.191 mmol) in DMF (1mL) were sequentially added N,N-diisopropylethylamine (0.100 mL, 0.574mmol),(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,HCl (0.072 g, 0.191 mmol) and BOP (0.093 g, 0.211 mmol) at roomtemperature. The contents were stirred at room temperature for 60 hours.The reaction mixture was subjected to preparative HPLC (Phenomex LunaAXIA 30×100 mm; 10 min. gradient; solvent A=10% MeOH, 90% H₂O, 0.1% TFA,solvent B=90% MeOH, 10% H₂O, 0.1% TFA) and fractions corresponding tothe desired products were isolated and concentrated. The residue waspartitioned between dichloromethane (20 mL) and saturated aqueous sodiumbicarbonate solution (3 mL). The dichloromethane layer was dried oversodium sulfate and concentrated to yield a solid. This solid wassubjected to further preparative HPLC separation under the followingconditions (Column: Princeton CN, 250×3 cm, 5 μm; Flow rate: 150 mL/min;Mobile Phase: CO₂/MeOH+0.1% TEA=88/12; Inj. Vol.=0.5 mL, Wavelength=220nM; peak 1, retention time=3.83 min; peak 2, retention time=3.98 min;peak 3, retention time=4.11 min) Peaks 1 and 2 correspond to the chlorocompound. Peak 3 corresponds to the deschloro compound. Each fractionwas individually concentrated (peak 1: 0.014 g, 0.027 mmol, 13.94%yield; peak 2: 0.024 g, 0.046 mmol, 23.89% yield; peak 3: 0.014 g, 0.029mmol, 14.92% yield).

Peak 1: Anal. RP-HPLC: Retention time=2.86 min (YMC S5 Combi ODS 4.6×50mm; 4 min. gradient; solvent A=10% MeOH, 90% H₂O, 0.2% H₃PO₄, solventB=90% MeOH, 10% H₂O, 0.2% H₃PO₄). Purity: >95%.; MS (ESI) m/z 524.29(M+H)⁺Peak 2: Anal. RP-HPLC: Retention time=2.82 min (YMC S5 Combi ODS 4.6×50mm; 4 min. gradient; solvent A=10% MeOH, 90% H₂O, 0.2% H₃PO₄, solventB=90% MeOH, 10% H₂O, 0.2% H₃PO₄. Purity: >95%.; MS (ESI) m/z 524.30(M+H)⁺Peak 3: Anal. RP-HPLC: Retention time=2.82 min (YMC S5 Combi ODS 4.6×50mm; 4 min. gradient; solvent A=10% MeOH, 90% H₂O, 0.2% H₃PO₄, solventB=90% MeOH, 10% H₂O, 0.2% H₃PO₄. Purity: >95%.; MS (ESI) m/z 490.30(M+H)⁺

Example 10(7R)-1-acetyl-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide

To a solution of(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide(from experiment 9, step C, peak 3, 13 mg, 0.027 mmol) indichloromethane (0.5 mL) was added triethylamine (7.39 μL, 0.053 mmol)and acetic anhydride (4.06 mg, 0.040 mmol). The resulting mixture wasstirred at room temperature for 2 hours. The reaction mixture wassubjected to preparative HPLC (Shimadzu VP-ODS 20×100 mm, 8 min.gradient, Solvent A: 10% MeOH, 90% H₂O, 0.1% TFA; Solvent B: 90% MeOH,10% H₂O, 0.1% TFA). The peak corresponding to the desired product wascollected, concentrated and freeze dried to give(7R)-1-acetyl-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide(6 mg, 0.011 mmol, 42.5% yield) as a solid. Anal. RP-HPLC: Retentiontime=3.44 min (YMC S5 ODS 4.6×50 mm; gradient solvent A: 10% MeOH, 90%H₂O, 0.2% H₃PO₄; Solvent B: 90% MeOH, 10% H2O, 0.2% H₃PO₄).Purity: >95%; MS (ESI) m/z 532 M⁺

Example 11N—((R)-1-((S)-4-(4-Chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-oxaspiro[2.4]heptane-5-carboxamide

Step A: Ethyl 3-methylenecyclopentanecarboxylate

The following reaction was performed in a 15 mL sealed tube, behind asafety shield. To 2-((trimethylsilyl)methyl)allyl acetate (2.227 mL,10.73 mmol) in toluene (2 mL) was sequentially added ethyl acrylate(2.92 mL, 26.8 mmol), triisopropyl phosphite (1.474 mL, 6.44 mmol) andpalladium(II)acetate (0.241 g, 1.073 mmol). The contents were purgedwith nitrogen gas for 5 min. and heated at 100° C. for 45 h. Thereaction mixture was cooled to RT and concentrated under reducedpressure. The liquid was transferred to a 50 mL round bottom flask anddistilled using a high vacuum pump. Fractions distilling at 50° C. werecollected and concentrated to yield 1.317 g (80%) of ethyl3-methylenecyclopentanecarboxylate. ¹H NMR (400 MHz, CDCl₃) δ ppm4.90-4.85 (m, 2H), 4.14 (q, 2H, J=7.0 Hz), 2.87-2.78 (m, 1H), 2.61-2.53(m, 2H), 2.51-2.23 (m, 2H), 2.09-1.82 (m, 2H), 1.29-1.23 (t, 3H, J=7.2Hz).

Step B: Lithium 3-methylenecyclopentanecarboxylate

A mixture of ethyl 3-methylenecyclopentanecarboxylate (900 mg, 5.84mmol) and lithium hydroxide (280 mg, 11.67 mmol) in THF (15 mL) andwater (6 mL) was heated at 65° C. for 18 hours. Analysis by TLCindicated that the starting ester had been completely consumed. Theorganic solvents were removed via rotary evaporator, and the remainingaqueous solution was freeze-dried to yield lithium3-methylenecyclopentanecarboxylate. ¹H NMR (400 MHz, CD₃OD) δ ppm4.84-4.73 (m, 2H), 2.71-2.59 (m, 1H), 2.57-2.35 (m, 3H), 2.29-2.16 (m,1H), 2.01-1.91 (1H, m), 1.84-1.72 (m, 1H,).

Step C:N—((R)-1-((S)-4-(4-Chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-methylenecyclopentanecarboxamide

A mixture of lithium 3-methylenecyclopentanecarboxylate (0.770 g, 5.83mmol), and Hunig's base (3.56 mL, 20.40 mmol) in DMF (10 mL) was treatedwith(R)-2-amino-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methylbutan-1-one,HCl (2.188 g, 5.83 mmol), then BOP (2.84 g, 6.41 mmol), and the mixturewas stirred at room temperature for 2 hours. The reaction mixture waspoured into 85:15 ethyl acetate/hexanes (200 mL), and the cloudysolution was washed 3× with water, and once with brine. The organicphase was dried over sodium sulfate and concentrated in vacuo. Theresidue was purified via MPLC over a 40 g silica gel column, eluting at40 mL/min with 20% then 55% ethyl acetate/hexanes to yieldN—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-methylenecyclopentanecarboxamide(1.89 g, 4.23 mmol, 72.5% yield) as a glass. ¹H NMR (400 MHz, CD₃OD)(NMR shows several rotomers) δ ppm 7.49 (d, 1H, J=8.8 Hz), 7.46 (d, 1H,J=8.4 Hz), 7.34-7.27 (m, 2H), 4.86-4.82 (m, 2H+H₂O), 4.60-4.51 (m,0.7H), 4.13-4.00 (m, 0.4H+residual ethyl acetate), 3.66-3.55 (m, 1H),3.52 (d, 0.6; H, J=12.7 Hz), 3.17 (td, 0.6; H, J=13.1, 2.9 Hz), 3.11 (d,0.5; H, J=12.3 Hz), 2.90-2.78 (m, 1H), 2.70 (td, 0.5; H, J=13.6, 4.8Hz), 2.61 (td, 0.7; H, J=13.5, 5.1 Hz), 2.56-2.40 (m, 3H), 2.39-2.22 (m,1H), 2.16 (dddd, 0.5; H, J=13.7, 6.8, 6.7, 6.6 Hz), 2.09-1.67 (m,2.7H+residual ethyl acetate), 1.60 (d, 0.5; H, J=14.1 Hz), 1.51 (d, 0.5;H, J=13.6 Hz), 1.32 (d, 0.6; H, J=5.7 Hz), 1.04 (d, 1.4; H, J=6.6 Hz),0.99 (dd, 1.4; H, J=6.8, 1.1 Hz), 0.92 (d, 3.2; H, J=6.6 Hz), 0.81 (d,3H, J=4.8 Hz), 0.75 (d, 3H, J=4.8 Hz).

LCMS Method:

Injection Volume: 10 μL; Start % B=50; Final % B=100

Gradient Time=4 minutes; Flow Rate=4 mL/minute; Wavelength=220 nM

Solvent A=10:90 methanol:water+0.1% TFA; Solvent B=90:10methanol:water+0.1% TFA; Column=Waters Sunfire C18 4.6×50 mm

Retention Time=3.10 minutes

MS (ESI⁺)=448.1 (M+H)⁺.

Step D Example 11

A mixture ofN—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3-methylenecyclopentanecarboxamide(535 mg, 1.197 mmol) and sodium bicarbonate (141 mg, 1.676 mmol) inmethylene chloride (4 mL) and water (2 mL) was cooled to 5° C. andtreated with m-CPBA (318 mg, 1.436 mmol). The mixture was stirred at 5°C. for 1 hour, then allowed to warm to room temperature and stirred for18 hours. The mixture was cooled to 5° C., treated with saturated sodiumsulfite, and stirred at room temperature for 30 minutes. The layers wereseparated, and the aqueous phase was extracted 3× with methylenechloride. The organic phases were combined and concentrated in vacuo,and the residue was taken up in ethyl acetate. The solution was washed3× with saturated sodium carbonate, once with water, and once withbrine, then dried over sodium sulfate, and concentrated in vacuo toyield the title compound (535 mg, 1.155 mmol, 97% yield) as a glass anda mixture of diastereomers. ¹H NMR (400 MHz, CD₃OD) (NMR shows severalrotomers) δ ppm 7.54-7.40 (m, 2H), 7.37-7.26 (m, 2H), 4.90-4.82 (m,1H+H₂O), 4.63-4.48 (m, 0.6H), 4.12-3.96 (m, 1H+residual ethyl acetate),3.68-3.46 (m, 1.6H), 3.23-3.02 (m, 1.6H), 2.98-2.90 (m, 0.4H), 2.90-2.77(m, 2H), 2.76-2.53 (m, 1H), 2.31-1.79 (m, 6H+residual ethyl acetate),1.79-1.64 (m, 1.6H), 1.60 (bd, 0.4; H, J=14.1 Hz), 1.57-1.48 (m, 0.6H),1.04 (dd, 1.4; H, J=6.6, 3.1 Hz), 1.00 (t, 1.6; H, J=6.4 Hz), 0.96-0.90(m, 3.2H), 0.87-0.79 (dd, 3H, J=5.5, 3.0 Hz), 0.79-0.71 (m, 3H).

LCMS Method:

Start % B=0; Final % B=95; Gradient Time=2 minutes

Flow Rate=4 mL/minute; Wavelength=220 nM

Solvent A=10:90 acetonitrile:water+NH₄OAc; Solvent B=90:10acetonitrile:water+NH₄OAc; Column=Phenomenex Luna C18 4.6×30 mm 3μ

Retention Time=0.92 minutes

MS (ESI⁺)=463.1 (M+H)⁺.

Example 12N—((R)-1-((S)-4-(4-Chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-oxaspiro[3.4]octane-6-carboxamide

A suspension of trimethylsulfoxonium iodide (67.8 mg, 0.308 mmol) intert-butanol (2 mL) was treated with potassium tert-butoxide (34.6 mg,0.308 mmol). The mixture was heated at 50° C. for 1 hour, then treatedwithN—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-oxaspiro[2.4]heptane-5-carboxamide(46 mg, 0.099 mmol), and the reaction was stirred at 50° C. for 2 days.The reaction was quenched with saturated ammonium chloride (1 mL), andthe mixture was poured into ethyl acetate (15 mL). The layers wereseparated, and the organic phase was washed 3× with water and once withbrine, then dried over sodium sulfate and concentrated in vacuo. Thecrude material was purified via preparative LC/MS with the followingconditions: Column. Waters SunFire C18, 19×150 mm, 5-μm particles; GuardColumn. None; Mobile Phase A: 5:95 acetonitrile:water with 10-mMammonium acetate; Mobile Phase B: 95:5 acetonitrile:water with 10-mMammonium acetate; Gradient: 15-100% B over 15 minutes, then a 5-minutehold at 100% B; Flow: 20 mL/min. Fractions containing the desiredproduct were combined and dried via centrifugal evaporation. The yieldof the desired product as a diastereomeric mixture was 15.4 mg, and itspurity was 96%. Two analytical LC/MS injections were used to determinethe final purity. Injection 1 conditions: Column: Supelco AscentisExpress C18, 4.6×50 mm, 2.7-μm particles; Mobile Phase A: 5:95acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5acetonitrile:water with 10 mM ammonium acetate; Temperature: 45° C.;Gradient: 0-100% B over 5.3 minutes, then a 1.7-minute hold at 100% B;Flow: 3 mL/min. Retention time=2.91. Injection 2 conditions: Column:Waters XBridge C18, 4.6×50 mm, 2.5-1 μm particles; Mobile Phase A: 5:95acetonitrile:water with 0.1% ammonium hydroxide; Mobile Phase B: 95:5acetonitrile:water with 0.1% ammonium hydroxide; Temperature: 45° C.;Gradient: 0-100% B over 7 minutes, then a 1-minute hold at 100% B; Flow:3 mL/min. Retention time=3.79 minutes. Proton NMR was acquired indeuterated methanol. MS (ESI⁺)=476.9 (M+H)⁺. ¹H NMR (400 MHz, CD₃OD)(NMR shows several rotomers) δ ppm 7.53-7.42 (m, 2H), 7.36-7.26 (m, 2H),4.83-4.76 (m, 1H), 4.58-4.42 (m, 2H), 4.13-3.98 (m, 1H), 3.66-3.58 (bd,1H, J=13.3 Hz), 3.58-3.46 (m, 1H), 3.22-3.06 (m, 1H), 2.95-2.86 (m,0.4H), 2.83-2.56 (m, 3.2H), 2.38-1.69 (m, 7H), 1.59 (d, 0.5; H, J=14.3Hz), 1.51 (d, 0.5; H, J=14.6 Hz), 1.47-1.42 (m, 0.2H), 1.04 (d, 1.5; H,J=6.5 Hz), 0.98 (t, 1.5; H, J=6.2 Hz), 0.92 (d, 3H, J=5.3 Hz), 0.81 (d,3H, J=4.8 Hz), 0.75 (br. s, 3H.).

Utility

In general, compounds of the present invention, such as particularcompounds disclosed in the preceding examples, have been shown to bemodulators of chemokine receptor activity (for example, by displaying Kivalues<10,000 nM in a binding assay such as those set forth below). Bydisplaying activity as modulators of chemokine receptor activity,compounds of the present invention are expected to be useful in thetreatment of human diseases associated with chemokines and their cognatereceptors.

CCR1 Ligand Binding Scintillation Proximity Assay (SPA)

For radioligand competition studies, a final concentration of 1×10⁵THP-1 monocytic leukemia cells are combined with 100 μg of LS WGA PSbeads (Amersham, Cat.#: RPNQ 0260) in 40 μl of assay buffer (RPMI 1640without phenol red, 50 mM HEPES, 5 mM MgCl₂, 1 mM CaCl₂, 0.1% BSA). TheTHP-1 cell/bead mixture is added to each well of a 384-well assay plate(PerkinElmer, Cat. #:6007899) containing test compound in 3-fold serialdilution, with final concentrations ranging from 8 μM to 140 pM. A finalconcentration of 0.1 nM [¹²⁵I]-MIP-1α (PerkinElmer, Cat. #NEX298) in 20μl assay buffer is added to the reaction. Sealed assay plates areincubated at room temperature for 12 h then analyzed by LEADseeker™.

The competition data of compound (I) over a range of concentrations isplotted as percentage inhibition of radioligand specific bound in theabsence of test compound (percent of total signal). After correcting fornon-specific binding, IC₅₀ values are determined. The IC₅₀ value isdefined as the concentration of test compound needed to reduce[¹²⁵I]-MIP-1α specific binding by 50% and is calculated using the fourparameter logistic equation to fit the normalized data. The Ki valuesare determined by application of the Cheng-Prusoff equation to the IC₅₀values, where K_(i)=IC₅₀/(1+ligand concentration/K_(d)) The Kd of[¹²⁵I]-MIP-1α in THP-1 cells is 0.1 nM. Each experiment is run induplicate.

Compounds of the present invention were tested in the assay describedabove and the results are shown in Table 1.

TABLE 1 Example CCR1 Binding Ki 3-Isomer 2 2.4 nM 5-Isomer 2 6.6 nM9-Peak 3 193 nM 

Mammalian chemokine receptors provide a target for interfering with orpromoting immune cell function in a mammal, such as a human. Compoundsthat inhibit or promote chemokine receptor function are particularlyuseful for modulating immune cell function for therapeutic purposes.

Accordingly, the present invention is directed to compounds which areuseful in the prevention and/or treatment of a wide variety ofinflammatory, infectious, and immunoregulatory disorders and diseases,including asthma and allergic diseases, infection by pathogenic microbes(which, by definition, includes viruses), as well as autoimmunepathologies such as the rheumatoid arthritis and atherosclerosis.

For example, an instant compound which inhibits one or more functions ofa mammalian chemokine receptor (e.g., a human chemokine receptor) may beadministered to inhibit (i.e., reduce or prevent) inflammation orinfectious disease. As a result, one or more inflammatory process, suchas leukocyte emigration, adhesion, chemotaxis, exocytosis (e.g., ofenzymes, histamine) or inflammatory mediator release, is inhibited.

Similarly, an instant compound which promotes one or more functions ofthe mammalian chemokine receptor (e.g., a human chemokine) asadministered to stimulate (induce or enhance) an immune or inflammatoryresponse, such as leukocyte emigration, adhesion, chemotaxis, exocytosis(e.g., of enzymes, histamine) or inflammatory mediator release,resulting in the beneficial stimulation of inflammatory processes. Forexample, eosinophils can be recruited to combat parasitic infections. Inaddition, treatment of the aforementioned inflammatory, allergic andautoimmune diseases can also be contemplated for an instant compoundwhich promotes one or more functions of the mammalian chemokine receptorif one contemplates the delivery of sufficient compound to cause theloss of receptor expression on cells through the induction of chemokinereceptor internalization or the delivery of compound in a manner thatresults in the misdirection of the migration of cells.

In addition to primates, such as humans, a variety of other mammals canbe treated according to the method of the present invention. Forinstance, mammals, including but not limited to, cows, sheep, goats,horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine,canine, feline, rodent or murine species can be treated. However, themethod can also be practiced in other species, such as avian species.The subject treated in the methods above is a mammal, male or female, inwhom modulation of chemokine receptor activity is desired. “Modulation”as used herein is intended to encompass antagonism, agonism, partialantagonism and/or partial agonism.

Diseases or conditions of human or other species which can be treatedwith inhibitors of chemokine receptor function, include, but are notlimited to: inflammatory or allergic diseases and conditions, includingrespiratory allergic diseases such as asthma, allergic rhinitis,hypersensitivity lung diseases, hypersensitivity pneumonitis,eosinophilic cellulitis (e.g., Well's syndrome), eosinophilic pneumonias(e.g., Loeffler's syndrome, chronic eosinophilic pneumonia),eosinophilic fasciitis (e.g., Shulman's syndrome), delayed-typehypersensitivity, interstitial lung diseases (ILD) (e.g., idiopathicpulmonary fibrosis, or ILD associated with rheumatoid arthritis,systemic lupus erythematosus, ankylosing spondylitis, systemicsclerosis, Sjorgren's syndrome, polymyositis or dermatomyositis);systemic anaphylaxis or hypersensitivity responses, drug allergies(e.g., to penicillin, cephalosporins), eosinophilia-myalgia syndrome dueto the ingestion of contaminated tryptophan, insect sting allergies;autoimmune diseases, such as rheumatoid arthritis, psoriatic arthritis,multiple sclerosis, systemic lupus erythematosus, myasthenia gravis,juvenile onset diabetes; glomerulonephritis, autoimmune thyroiditis,Behcet's disease; graft rejection (e.g., in transplantation), includingallograft rejection or graft-versus-host disease; inflammatory boweldiseases, such as Crohn's disease and ulcerative colitis;spondyloarthropathies; scleroderma; psoriasis (including T-cell mediatedpsoriasis) and inflammatory dermatoses such as an dermatitis, eczema,atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis(e.g., necrotizing, cutaneous, and hypersensitivity vasculitis);eosinophilic myositis, eosinophilic fasciitis; cancers with leukocyteinfiltration of the skin or organs. Other diseases or conditions inwhich undesirable inflammatory responses are to be inhibited can betreated, including, but not limited to, reperfusion injury,atherosclerosis, certain hematological malignancies, cytokine-inducedtoxicity (e.g., septic shock, endotoxic shock), polymyositis,dermatomyositis. Infectious diseases or conditions of human or otherspecies which can be treated with inhibitors of chemokine receptorfunction, include, but are not limited to, HIV.

Diseases or conditions of humans or other species which can be treatedwith promoters of chemokine receptor function, include, but are notlimited to: immunosuppression, such as that in individuals withimmunodeficiency syndromes such as AIDS or other viral infections,individuals undergoing radiation therapy, chemotherapy, therapy forautoimmune disease or drug therapy (e.g., corticosteroid therapy), whichcauses immunosuppression; immunosuppression due to congenital deficiencyin receptor function or other causes; and infections diseases, such asparasitic diseases, including, but not limited to helminth infections,such as nematodes (round worms); (Trichuriasis, Enterobiasis,Ascariasis, Hookworm, Strongyloidiasis, Trichinosis, filariasis);trematodes (flukes) (Schistosomiasis, Clonorchiasis), cestodes (tapeworms) (Echinococcosis, Taeniasis saginata, Cysticercosis); visceralworms, visceral larva migraines (e.g., Toxocara), eosinophilicgastroenteritis (e.g., Anisaki sp., Phocanema sp.), cutaneous larvamigraines (Ancylostona braziliense, Ancylostoma caninum). The compoundsof the present invention are accordingly useful in the prevention andtreatment of a wide variety of inflammatory, infectious andimmunoregulatory disorders and diseases.

In addition, treatment of the aforementioned inflammatory, allergic andautoimmune diseases can also be contemplated for promoters of chemokinereceptor function if one contemplates the delivery of sufficientcompound to cause the loss of receptor expression on cells through theinduction of chemokine receptor internalization or delivery of compoundin a manner that results in the misdirection of the migration of cells.

In another aspect, the instant invention may be used to evaluate theputative specific agonists or antagonists of a G protein coupledreceptor. The present invention is directed to the use of thesecompounds in the preparation and execution of screening assays forcompounds that modulate the activity of chemokine receptors.Furthermore, the compounds of this invention are useful in establishingor determining the binding site of other compounds to chemokinereceptors, e.g., by competitive inhibition or as a reference in an assayto compare its known activity to a compound with an unknown activity.When developing new assays or protocols, compounds according to thepresent invention could be used to test their effectiveness.Specifically, such compounds may be provided in a commercial kit, forexample, for use in pharmaceutical research involving the aforementioneddiseases. The compounds of the instant invention are also useful for theevaluation of putative specific modulators of the chemokine receptors.In addition, one could utilize compounds of this invention to examinethe specificity of G protein coupled receptors that are not thought tobe chemokine receptors, either by serving as examples of compounds whichdo not bind or as structural variants of compounds active on thesereceptors which may help define specific sites of interaction.

The compounds of the present invention are used to treat or preventdisorders selected from rheumatoid arthritis, osteoarthritis, septicshock, atherosclerosis, aneurysm, fever, cardiovascular effects,haemodynamic shock, sepsis syndrome, post ischemic reperfusion injury,malaria, Crohn's disease, inflammatory bowel diseases, mycobacterialinfection, meningitis, psoriasis, congestive heart failure, fibroticdiseases, cachexia, graft rejection, autoimmune diseases, skininflammatory diseases, multiple sclerosis, radiation damage, hyperoxicalveolar injury, HIV, HIV dementia, non-insulin dependent diabetesmellitus, asthma, allergic rhinitis, atopic dermatitis, idiopathicpulmonary fibrosis, bullous pemphigoid, helminthic parasitic infections,allergic colitis, eczema, conjunctivitis, transplantation, familialeosinophilia, eosinophilic cellulitis, eosinophilic pneumonias,eosinophilic fasciitis, eosinophilic gastroenteritis, drug inducedeosinophilia, cystic fibrosis, Churg-Strauss syndrome, lymphoma,Hodgkin's disease, colonic carcinoma, Felty's syndrome, sarcoidosis,uveitis, Alzheimer, Glomerulonephritis, and systemic lupuserythematosus.

In another aspect, the compounds are used to treat or preventinflammatory disorders selected from rheumatoid arthritis,osteoarthritis, atherosclerosis, aneurysm, fever, cardiovasculareffects, Crohn's disease, inflammatory bowel diseases, psoriasis,congestive heart failure, multiple sclerosis, autoimmune diseases, skininflammatory diseases.

In another aspect, the compounds are used to treat or preventinflammatory disorders selected from rheumatoid arthritis,osteoarthritis, atherosclerosis, Crohn's disease, inflammatory boweldiseases, and multiple sclerosis.

Combined therapy to prevent and treat inflammatory, infectious andimmunoregulatory disorders and diseases, including asthma and allergicdiseases, as well as autoimmune pathologies such as rheumatoid arthritisand atherosclerosis, and those pathologies noted above is illustrated bythe combination of the compounds of this invention and other compoundswhich are known for such utilities. For example, in the treatment orprevention of inflammation, the present compounds may be used inconjunction with an anti-inflammatory or analgesic agent such as anopiate agonist, a lipoxygenase inhibitor, a cyclooxygenase-2 inhibitor,an interleukin inhibitor, such as an interleukin-1 inhibitor, a tumornecrosis factor inhibitor, an NMDA antagonist, an inhibitor nitric oxideor an inhibitor of the synthesis of nitric oxide, a non-steroidalanti-inflammatory agent, a phosphodiesterase inhibitor, or acytokine-suppressing anti-inflammatory agent, for example with acompound such as acetaminophen, aspirin, codeine, fentaynl, ibuprofen,indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, asteroidal analgesic, sufentanyl, sunlindac, interferon alpha and thelike. Similarly, the instant compounds may be administered with a painreliever; a potentiator such as caffeine, an H2-antagonist, simethicone,aluminum or magnesium hydroxide; a decongestant such as phenylephrine,phenylpropanolamine, pseudophedrine, oxymetazoline, ephinephrine,naphazoline, xylometazoline, propylhexedrine, or levodesoxy-ephedrine;and antitussive such as codeine, hydrocodone, caramiphen,carbetapentane, or dextramethorphan; a diuretic; and a sedating ornon-sedating antihistamine. Likewise, compounds of the present inventionmay be used in combination with other drugs that are used in thetreatment/prevention/suppression or amelioration of the diseases orconditions for which compound of the present invention are useful. Suchother drugs may be administered, by a route and in an amount commonlyused therefore, contemporaneously or sequentially with a compound of thepresent invention. When a compound of the present invention is usedcontemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compound ofthe present invention may be used. Accordingly, the pharmaceuticalcompositions of the present invention include those that also containone or more other active ingredients, in addition to a compound of thepresent invention.

Examples of other active ingredients that may be combined with acompound of the present invention, either administered separately or inthe same pharmaceutical compositions, include, but are not limited to:(a) integrin antagonists such as those for selectins, ICAMs and VLA-4;(b) steroids such as beclomethasone, methylprednisolone, betamethasone,prednisone, dexamethasone, and hydrocortisone; (c) immunosuppressantssuch as cyclosporin, tacrolimus, rapamycin and other FK-506 typeimmunosuppressants; (d) antihistamines (H1-histamine antagonists) suchas bromopheniramine, chlorpheniramine, dexchlorpheniramine,triprolidine, clemastine, diphenhydramine, diphenylpyraline,tripelennamine, hydroxyzine, methdilazine, promethazine, trimeprazine,azatadine, cyproheptadine, antazoline, pheniramine pyrilamine,astemizole, terfenadine, loratadine, cetirizine, fexofenadine,descarboethoxyloratadine, and the like; (e) non-steroidalanti-asthmatics such as b2-agonists (terbutaline, metaproterenol,fenoterol, isoetharine, albuteral, bitolterol, and pirbuterol),theophylline, cromolyn sodium, atropine, ipratropium bromide,leukotriene antagonists (zafirlukast, montelukast, pranlukast,iralukast, pobilukast, SKB-102,203), leukotriene biosynthesis inhibitors(zileuton, BAY-1005); (f) non-steroidal anti-inflammatory agents(NSAIDs) such as propionic acid derivatives (alminoprofen, benxaprofen,bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen,ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin,pirprofen, pranoprofen, suprofen, tiaprofenic acid, and tioxaprofen),acetic acid derivatives (indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin, andzomepirac), fenamic acid derivatives (flufenamic acid, meclofenamicacid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxicams(isoxicam, piroxicam, sudoxicam and tenoxican), salicylates (acetylsalicylic acid, sulfasalazine) and the pyrazolones (apazone,bezpiperylon, feprazone, mofebutazone, oxyphenbutazone, phenylbutazone);(g) cyclooxygenase-2 (COX-2) inhibitors; (h) inhibitors ofphosphodiesterase type IV (PDE-IV); (i) other antagonists of thechemokine receptors; (j) cholesterol lowering agents such as HMG-COAreductase inhibitors (lovastatin, simvastatin and pravastatin,fluvastatin, atorvastatin, and other statins), sequestrants(cholestyramine and colestipol), nicotonic acid, fenofibric acidderivatives (gemfibrozil, clofibrat, fenofibrate and benzafibrate), andprobucol; (k) anti-diabetic agents such as insulin, sulfonylureas,biguanides (metformin), a-glucosidase inhibitors (acarbose) andglitazones (troglitazone and pioglitazone); (l) preparations ofinterferons (interferon alpha-2a, interferon-2B, interferon alpha-N3,interferon beta-1a, interferon beta-1b, interferon gamma-1b); (m)antiviral compounds such as efavirenz, nevirapine, indinavir,ganciclovir, lamivudine, famciclovir, and zalcitabine; (n) othercompound such as 5-aminosalicylic acid an prodrugs thereof,anti-metabolites such as azathioprine and 6-mercaptopurine, andcytotoxic cancer chemotherapeutic agents. The weight ratio of thecompound of the present invention to the second active ingredient may bevaried and will depend upon the effective doses of each ingredient.

Generally, an effective dose of each will be used. Thus, for example,when a compound of the present invention is combined with an NSAID theweight ratio of the compound of the present invention to the NSAID willgenerally range from about 1000:1 to about 1:1000, or alternatively fromabout 200:1 to about 1:200. Combinations of a compound of the presentinvention and other active ingredients will generally also be within theaforementioned range, but in each case, an effective dose of each activeingredient should be used.

The compounds are administered to a mammal in a therapeuticallyeffective amount. By “therapeutically effective amount” it is meant anamount of a compound of Formula I that, when administered alone or incombination with an additional therapeutic agent to a mammal, iseffective to prevent or ameliorate the thromboembolic disease conditionor the progression of the disease.

Dosage and Formulation

The compounds of this invention can be administered in such oral dosageforms as tablets, capsules (each of which includes sustained release ortimed release formulations), pills, powders, granules, elixirs,tinctures, suspensions, syrups, and emulsions. They may also beadministered in intravenous (bolus or infusion), intraperitoneal,subcutaneous, or intramuscular form, all using dosage forms well knownto those of ordinary skill in the pharmaceutical arts. They can beadministered alone, but generally will be administered with apharmaceutical carrier selected on the basis of the chosen route ofadministration and standard pharmaceutical practice.

The dosage regimen for the compounds of the present invention will, ofcourse, vary depending upon known factors, such as the pharmacodynamiccharacteristics of the particular agent and its mode and route ofadministration; the species, age, sex, health, medical condition, andweight of the recipient; the nature and extent of the symptoms; the kindof concurrent treatment; the frequency of treatment; the route ofadministration, the renal and hepatic function of the patient, and theeffect desired. A physician or veterinarian can determine and prescribethe effective amount of the drug required to prevent, counter, or arrestthe progress of the thromboembolic disorder.

By way of general guidance, the daily oral dosage of each activeingredient, when used for the indicated effects, will range betweenabout 0.001 to 1000 mg/kg of body weight, or between about 0.01 to 100mg/kg of body weight per day, or alternatively, between about 1.0 to 20mg/kg/day. Intravenously, the doses will range from about 1 to about 10mg/kg/minute during a constant rate infusion. Compounds of thisinvention may be administered in a single daily dose, or the total dailydosage may be administered in divided doses of two, three, or four timesdaily.

Compounds of this invention can be administered in intranasal form viatopical use of suitable intranasal vehicles, or via transdermal routes,using transdermal skin patches. When administered in the form of atransdermal delivery system, the dosage administration will, of course,be continuous rather than intermittent throughout the dosage regimen.

The compounds are typically administered in admixture with suitablepharmaceutical diluents, excipients, or carriers (collectively referredto herein as pharmaceutical carriers) suitably selected with respect tothe intended form of administration, that is, oral tablets, capsules,elixirs, syrups and the like, and consistent with conventionalpharmaceutical practices.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic, pharmaceutically acceptable, inert carrier such as lactose,starch, sucrose, glucose, methyl cellulose, magnesium stearate,dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like;for oral administration in liquid form, the oral drug components can becombined with any oral, non-toxic, pharmaceutically acceptable inertcarrier such as ethanol, glycerol, water, and the like. Moreover, whendesired or necessary, suitable binders, lubricants, disintegratingagents, and coloring agents can also be incorporated into the mixture.Suitable binders include starch, gelatin, natural sugars such as glucoseor beta-lactose, corn sweeteners, natural and synthetic gums such asacacia, tragacanth, or sodium alginate, carboxymethylcellulose,polyethylene glycol, waxes, and the like. Lubricants used in thesedosage forms include sodium oleate, sodium stearate, magnesium stearate,sodium benzoate, sodium acetate, sodium chloride, and the like.Disintegrators include, without limitation, starch, methyl cellulose,agar, bentonite, xanthan gum, and the like.

The compounds of the present invention can also be administered in theform of liposome delivery systems, such as small unilamellar vesicles,large unilamellar vesicles, and multilamellar vesicles. Liposomes can beformed from a variety of phospholipids, such as cholesterol,stearylamine, or phosphatidylcholines.

Compounds of the present invention may also be coupled with solublepolymers as targetable drug carriers. Such polymers can includepolyvinylpyrrolidone, pyran copolymer,polyhydroxypropylmethacrylamide-phenol,polyhydroxyethylaspartamidephenol, or polyethyleneoxide-polylysinesubstituted with palmitoyl residues. Furthermore, the compounds of thepresent invention may be coupled to a class of biodegradable polymersuseful in achieving controlled release of a drug, for example,polylactic acid, polyglycolic acid, copolymers of polylactic andpolyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, andcrosslinked or amphipathic block copolymers of hydrogels.

Dosage forms (pharmaceutical compositions) suitable for administrationmay contain from about 1 milligram to about 100 milligrams of activeingredient per dosage unit. In these pharmaceutical compositions theactive ingredient will ordinarily be present in an amount of about0.5-95% by weight based on the total weight of the composition.

Gelatin capsules may contain the active ingredient and powderedcarriers, such as lactose, starch, cellulose derivatives, magnesiumstearate, stearic acid, and the like. Similar diluents can be used tomake compressed tablets. Both tablets and capsules can be manufacturedas sustained release products to provide for continuous release ofmedication over a period of hours. Compressed tablets can be sugarcoated or film coated to mask any unpleasant taste and protect thetablet from the atmosphere, or enteric coated for selectivedisintegration in the gastrointestinal tract.

Liquid dosage forms for oral administration can contain coloring andflavoring to increase patient acceptance.

In general, water, a suitable oil, saline, aqueous dextrose (glucose),and related sugar solutions and glycols such as propylene glycol orpolyethylene glycols are suitable carriers for parenteral solutions.Solutions for parenteral administration may contain a water soluble saltof the active ingredient, suitable stabilizing agents, and if necessary,buffer substances. Antioxidizing agents such as sodium bisulfate, sodiumsulfite, or ascorbic acid, either alone or combined, are suitablestabilizing agents. Also used are citric acid and its salts and sodiumEDTA. In addition, parenteral solutions can contain preservatives, suchas benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.

Suitable pharmaceutical carriers are described in Remington'sPharmaceutical Sciences, Mack Publishing Company, a standard referencetext in this field.

Representative useful pharmaceutical dosage-forms for administration ofthe compounds of this invention can be illustrated as follows:

Capsules

A large number of unit capsules can be prepared by filling standardtwo-piece hard gelatin capsules each with 100 milligrams of powderedactive ingredient, 150 milligrams of lactose, 50 milligrams ofcellulose, and 6 milligrams magnesium stearate.

Soft Gelatin Capsules

A mixture of active ingredient in a digestible oil such as soybean oil,cottonseed oil or olive oil may be prepared and injected by means of apositive displacement pump into gelatin to form soft gelatin capsulescontaining 100 milligrams of the active ingredient. The capsules shouldbe washed and dried.

Tablets

Tablets may be prepared by conventional procedures so that the dosageunit is 100 milligrams of active ingredient, 0.2 milligrams of colloidalsilicon dioxide, 5 milligrams of magnesium stearate, 275 milligrams ofmicrocrystalline cellulose, 11 milligrams of starch and 98.8 milligramsof lactose. Appropriate coatings may be applied to increase palatabilityor delay absorption.

Injectable

A parenteral composition suitable for administration by injection may beprepared by stirring 1.5% by weight of active ingredient in 10% byvolume propylene glycol and water. The solution should be made isotonicwith sodium chloride and sterilized.

Suspension

An aqueous suspension can be prepared for oral administration so thateach 5 mL contain 100 mg of finely divided active ingredient, 200 mg ofsodium carboxymethyl cellulose, 5 mg of sodium benzoate, 1.0 g ofsorbitol solution, U.S.P., and 0.025 mL of vanillin.

Where the compounds of this invention are combined with otheranticoagulant agents, for example, a daily dosage may be about 0.1 to100 milligrams of the compound of Formula I and about 1 to 7.5milligrams of the second anticoagulant, per kilogram of patient bodyweight. For a tablet dosage form, the compounds of this inventiongenerally may be present in an amount of about 5 to 10 milligrams perdosage unit, and the second anti-coagulant in an amount of about 1 to 5milligrams per dosage unit.

Where two or more of the foregoing second therapeutic agents areadministered with the compound of Formula I, generally the amount ofeach component in a typical daily dosage and typical dosage form may bereduced relative to the usual dosage of the agent when administeredalone, in view of the additive or synergistic effect of the therapeuticagents when administered in combination. Particularly when provided as asingle dosage unit, the potential exists for a chemical interactionbetween the combined active ingredients. For this reason, when thecompound of Formula I and a second therapeutic agent are combined in asingle dosage unit they are formulated such that although the activeingredients are combined in a single dosage unit, the physical contactbetween the active ingredients is minimized (that is, reduced). Forexample, one active ingredient may be enteric coated. By enteric coatingone of the active ingredients, it is possible not only to minimize thecontact between the combined active ingredients, but also, it ispossible to control the release of one of these components in thegastrointestinal tract such that one of these components is not releasedin the stomach but rather is released in the intestines. One of theactive ingredients may also be coated with a material which effects asustained-release throughout the gastrointestinal tract and also servesto minimize physical contact between the combined active ingredients.Furthermore, the sustained-released component can be additionallyenteric coated such that the release of this component occurs only inthe intestine. Still another approach would involve the formulation of acombination product in which the one component is coated with asustained and/or enteric release polymer, and the other component isalso coated with a polymer such as a low viscosity grade ofhydroxypropyl methylcellulose (HPMC) or other appropriate materials asknown in the art, in order to further separate the active components.The polymer coating serves to form an additional barrier to interactionwith the other component.

These as well as other ways of minimizing contact between the componentsof combination products of the present invention, whether administeredin a single dosage form or administered in separate forms but at thesame time by the same manner, will be readily apparent to those skilledin the art, once armed with the present disclosure.

While the invention has been described in detail and with reference tospecific embodiments thereof, it will be apparent to one skilled in theart that various changes and modifications can be made therein withoutdeparting from the spirit and scope thereof.

What is claimed is:
 1. A compound of Formula (I):

or stereoisomers or pharmaceutically acceptable salt forms thereof,wherein: the dashed line represents an optional double bond; Ring A isan optionally substituted three to nine membered mono- or bicyclicheterocyclic ring; T is

W is —OH; Z is a O or S; R and R₁ are independently hydrogen, alkyl,halo, C═O or —COalkyl; R₂ is hydrogen, alkyl or cycloalkyl; R₃, at eachoccurrence, are independently hydrogen or alkyl; R₄, at each occurrence,are independently hydrogen, halo, hydroxy or alkyl; R₅, at eachoccurrence, is hydrogen, halo, alkyl or cycloalkyl; m, at eachoccurrence, is 0-2; and n, at each occurrence, is 0-2.
 2. A compound ofFormula (Ia):

wherein Ring A is an optionally substituted five to seven memberedheterocyclic ring; T is

W is —OH; R and R₁ are independently hydrogen, alkyl, halo, C═O or—COalkyl; R₂ is C₁-C₄alkyl or cycloalkyl; R₃, at each occurrence, areindependently hydrogen or alkyl; R₄, at each occurrence, areindependently hydroxy or C₁-C₄alkyl; R₅, at each occurrence, is ahalogen atom; m, at each occurrence, is 0-2; and n, at each occurrence,is 0-2.
 3. A compound of Formula (Ib):

wherein Ring A is an optionally substituted five to six memberedheterocyclic ring; R and R₁ are independently hydrogen, alkyl, halo, C═Oor —COalkyl; R₂ is C₁-C₄alkyl; R₃, at each occurrence, are independentlyhydrogen or alkyl; R₄, at each occurrence, are independently C₁-C₄alkyl;R₅, at each occurrence, is —Cl; m, at each occurrence, is 0-2; and n, ateach occurrence, is 0-2.
 4. The compound of claim 1, or a stereoisomeror pharmaceutically acceptable salt form thereof, wherein the compoundis selected from(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-8-oxa-6-azaspiro[4.5]decane-2-carboxamide,(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6,8-diazaspiro[4.5]decane-2-carboxamide,N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6-oxa-8-azaspiro[4.5]decane-1-carboxamide,(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-oxo-1-oxa-3-azaspiro[4.4]nonane-7-carboxamide;(2R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-7-oxo-6-oxa-8-azaspiro[4.5]decane-2-carboxamide,(3R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-3′-methyl-6′-oxo-2′,5′,6′,7′-tetrahydro-spiro[cyclopentane-1,4′-pyrazolo[3,4-b]pyridine]-3-carboxamide,(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxamide,(7R)—N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-methyl-2,4-dioxo-1,3-diazaspiro[4.4]nonane-7-carboxamide,(7R)-3-chloro-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide,(7R)-1-acetyl-N—((R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-azaspiro[4.4]nonane-7-carboxamide,N—((R)-1-((S)-4-(4-Chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-oxaspiro[2.4]heptane-5-carboxamide,andN—((R)-1-((S)-4-(4-Chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1-oxaspiro[3.4]octane-6-carboxamide.5. A pharmaceutical composition comprised of a pharmaceuticallyacceptable carrier and a therapeutically effective amount of one or morecompound of claim
 1. 6. A method for modulation of chemokine orchemokine receptor activity comprising administering to a patient inneed thereof a therapeutically effective amount of one or more compoundof claim
 1. 7. The method of claim 6, wherein the chemokine or chemokinereceptor activity is CCR-1 or CCR-1 receptor activity.
 8. A method fortreating a disorder comprising administering to a patient in needthereof a therapeutically effective amount of one or more compound ofclaim 1; wherein said disorder is selected from osteoarthritis,aneurysm, fever, cardiovascular effects, Crohn's disease, congestiveheart failure, autoimmune diseases, HIV-infection, HIV-associateddementia, psoriasis, idiopathic pulmonary fibrosis, transplantarteriosclerosis, physically- or chemically-induced brain trauma,inflammatory bowel disease, alveolitis, colitis, systemic lupuserythematosus, nephrotoxic serum nephritis, glomerularnephritis, asthma,multiple sclerosis, atherosclerosis, rheumatoid arthritis, restinosis,organ transplantation, and cancer.
 9. A method for treating inflammatorydiseases comprising administering to a patient in need thereof atherapeutically effective amount of one or more compound of claim
 1. 10.A method for treating inflammatory diseases which are at least partiallymediated by CCR-1 comprising administering to a patient in need thereofa therapeutically effective amount of one or more compound of claim 1.11. A method for preparing a medicament for the treatment ofosteoarthritis, aneurysm, fever, cardiovascular effects, Crohn'sdisease, congestive heart failure, autoimmune diseases, HIV-infection,HIV-associated dementia, psoriasis, idiopathic pulmonary fibrosis,transplant arteriosclerosis, physically- or chemically-induced braintrauma, inflammatory bowel disease, alveolitis, colitis, systemic lupuserythematosus, nephrotoxic serum nephritis, glomerularnephritis, asthma,multiple sclerosis, atherosclerosis, and rheumatoid arthritis comprisingformulating one or more compound of claim
 1. 12. A method for treating apatient in need of therapy comprising administering to said patient inneed thereof a therapeutically effective amount of one or more compoundof claim
 1. 13. A method for modulation of CCR-1 receptor activitycomprising administering to a patient in need thereof a therapeuticallyeffective amount of a composition of claim
 5. 14. A method for treatinga disorder comprising administering to a patient in need thereof atherapeutically effective amount of a composition of claim 5, whereinsaid disorder is selected from osteoarthritis, aneurysm, fever,cardiovascular effects, Crohn's disease, congestive heart failure,autoimmune diseases, HIV-infection, HIV-associated dementia, psoriasis,idiopathic pulmonary fibrosis, transplant arteriosclerosis, physically-or chemically-induced brain trauma, inflammatory bowel disease,alveolitis, colitis, systemic lupus erythematosus, nephrotoxic serumnephritis, glomerularnephritis, asthma, multiple sclerosis,atherosclerosis, rheumatoid arthritis, restinosis, organtransplantation, and cancer.
 15. A method for treating inflammatorydiseases comprising administering to a patient in need thereof atherapeutically effective amount of a composition of claim 5.